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<title>MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in Pseudomonas aeruginosa PAO1 - PubMed</title>
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<meta name="description" content="The C5-dicarboxylate α-ketoglutarate (α-KG) is a preferred nutrient source for the opportunistic pathogen &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt;. However, very little is known about how &lt;i&gt;P. aeruginosa&lt;/i&gt; detects and responds to α-KG in the environment. Our laboratory has previously shown that the MifS/Mif …"><meta name="keywords" content="pmid:32553056, PMC7654738, doi:10.1099/mic.0.000943, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov&#x27;t, Zaara Sarwar, Michael X Wang, Christopher T Nomura, Amino Acid Motifs, Amino Acid Sequence, Binding Sites, Biological Transport, Dicarboxylic Acids / metabolism, Escherichia coli / genetics, Gene Expression Regulation, Bacterial, Histidine Kinase / chemistry, Histidine Kinase / genetics, Histidine Kinase / metabolism*, Ketoglutaric Acids / metabolism*, Models, Molecular, Protein Domains, Protein Structure, Secondary, Pseudomonas aeruginosa / genetics, Pseudomonas aeruginosa / metabolism*, Signal Transduction, Substrate Specificity, Succinic Acid / metabolism, PubMed Abstract, NIH, NLM, NCBI, National Institutes of Health, National Center for Biotechnology Information, National Library of Medicine, MEDLINE"><meta name="robots" content="index,nofollow,noarchive"><meta property="og:title" content="MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in Pseudomonas aeruginosa PAO1 - PubMed"><meta property="og:url" content="https://pubmed.ncbi.nlm.nih.gov/32553056/"><meta property="og:description" content="The C5-dicarboxylate α-ketoglutarate (α-KG) is a preferred nutrient source for the opportunistic pathogen &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt;. However, very little is known about how &lt;i&gt;P. aeruginosa&lt;/i&gt; detects and responds to α-KG in the environment. Our laboratory has previously shown that the MifS/Mif …"><meta property="og:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:image:secure_url" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:type" content="website"><meta property="og:site_name" content="PubMed"><meta name="twitter:domain" content="pubmed.ncbi.nlm.nih.gov"><meta name="twitter:card" content="summary_large_image"><meta name="twitter:title" content="MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in Pseudomonas aeruginosa PAO1 - PubMed"><meta name="twitter:url" content="https://pubmed.ncbi.nlm.nih.gov/32553056/"><meta name="twitter:description" content="The C5-dicarboxylate α-ketoglutarate (α-KG) is a preferred nutrient source for the opportunistic pathogen &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt;. However, very little is known about how &lt;i&gt;P. aeruginosa&lt;/i&gt; detects and responds to α-KG in the environment. Our laboratory has previously shown that the MifS/Mif …"><meta name="twitter:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg">
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<input type="email" aria-label="Recipient Email Address" placeholder="email@example.com" id="email-to" class="email-to" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="100" required>
</div>
<div class="action-panel-control-wrap">
<label for="email-from" class="action-panel-label">
From:
</label>
<input type="email" aria-label="Sender Email Address" placeholder="email@example.com" id="email-from" class="email-from" pattern="^[a-zA-Z0-9_.+-]+@[a-zA-Z0-9-]+\.[a-zA-Z0-9-.]+$" maxlength="256">
</div>
<div class="action-panel-control-wrap">
<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="32553056">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="http://mic.microbiologyresearch.org/pubmed/content/journal/micro/10.1099/mic.0.000943"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3152&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=32553056&amp;db=pubmed&amp;nlmid=9430468"
title="See full text options at Ingenta plc"
data-ga-category="full_text"
data-ga-action="Ingenta plc"
data-ga-label="32553056"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.microbiologyresearch.org-marketing-MIC_linkout_logo.png" alt="Ingenta plc full text link"><span class="text">
Ingenta plc
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/32553056/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=32553056&amp;db=pubmed&amp;nlmid=9430468"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="32553056"
><span class="text">
Free PMC article
</span></a>
</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/32553056/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/32553056/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="32553056" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F32553056%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="VeALWOS1tjhUZaVkTAiNFrLBckQYPnkRc1kQzEln3U4oTB1awTVP9zhfKoyaIYSb"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
type="text"
name="add-to-new-collection"
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MifS, a DctB family histidine kinase, is a specific regulator of α-ketoglutarate response in <i>Pseudomonas aeruginosa</i> PAO1
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><sup class="key">4</sup> Center for Applied Microbiology, The State University of New York, College of Environmental Science and Forestry, Syracuse, New York, USA.</li>
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Abstract
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<p>
The C5-dicarboxylate α-ketoglutarate (α-KG) is a preferred nutrient source for the opportunistic pathogen <i>Pseudomonas aeruginosa</i>. However, very little is known about how <i>P. aeruginosa</i> detects and responds to α-KG in the environment. Our laboratory has previously shown that the MifS/MifR two-component signal transduction system regulates α-KG assimilation in <i>P. aeruginosa</i> PAO1. In an effort to better understand how this bacterium detects α-KG, we characterized the MifS sensor histidine kinase. In this study we show that although MifS is a homologue of the C4-dicarboxylate sensor DctB, it specifically responds to the C5-dicarboxylate α-KG. MifS activity increased &gt;10-fold in the presence of α-KG, while the related C5-dicarboxylate glutarate caused only a 2-fold increase in activity. All other dicarboxylates tested did not show any significant effect on MifS activity. Homology modelling of the MifS sensor domain revealed a substrate binding pocket for α-KG. Using protein modelling and mutational analysis, we identified nine residues that are important for α-KG response, including one residue that determines the substrate specificity of MifS. Further, we found that MifS has a novel cytoplasmic linker domain that is required for α-KG response and is probably involved in signal transduction from the sensor domain to the cytoplasmic transmitter domain. Until this study, DctB family histidine kinases were known to only respond to C4-dicarboxylates. Our work shows that MifS is a novel member of the DctB family histidine kinase that specifically responds to α-KG.
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Keywords:
</strong>
MifS; Pseudomonas aeruginosa; dicarboxylate sensor; sensor histidine kinase; two-component signal transduction systems; α-ketoglutarate.
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<p xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:p1="http://pubmed.gov/pub-one">The authors declare that there are no conflicts of interest.</p>
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Figures
</h2>
<div class="figures-list" id="slides-container"
itemscope itemtype="http://schema.org/ImageGallery">
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="0"
data-label-slug="fig-1">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/57702cb5622a/mic-166-867-g001.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-0"
role="button"
data-image-width="787"
data-image-height="483"
data-image-alt="Fig. 1."
data-pmc-id="PMC7654738"
data-figure-id="F1">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/c225dd3271e4/mic-166-867-g001.gif"
alt="Fig. 1." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="787">
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<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 1. </p>
</strong>
<div class="figure-caption-contents"><p> MifS is a DctB family… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 1. </p>
</strong>
<div class="figure-caption-contents"><p> MifS is a DctB family transmembrane histidine kinase. (a) Domain architecture of MifS… </p></div>
</div>
<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 1.
</strong>
<div class="figure-caption-contents">MifS is a DctB family transmembrane histidine kinase. (a) Domain architecture of MifS showing a periplasmic sensor domain (SD) flanked by two transmembrane helices (TMH1 and TMH2). TMH2 is followed by a cytoplasmic linker domain (LD) attached to a cytoplasmic transmitter domain (TD). (b) Alignment of the transmitter domain sequences of <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.2553">Pseudomonas aeruginosa</ext-link>
</i> MifS, COG4191 consensus sequence representing the DctB family of sensor histidine kinases, and HPK4 family histidine kinases from <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.2553">P. aeruginosa</ext-link>
</i> (<i>Pa</i>), <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3727">Myxococcus xanthus</ext-link>
</i> (<i>Mx</i>) and <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">Escherichia coli</ext-link>
</i> (<i>Ec</i>). The invariant residues of each transmitter domain conserved sequence box are shown in bold type. Conserved HPK4 sequence motifs are indicated with an underscore (_) . Sequence motifs conserved between MifS and DctB COG4191are highlighted in grey. Alignment was generated using <sc>clustal</sc> Omega [56] and displayed using ESPript 3.0 [57]. (c) Wild-type MifS, or a MifS variant harbouring a mutation in one of the essential transmitter domain residues (MifSH372A, MifSN483A, or MifSD517A), was expressed in <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the <i>mifR</i> gene and the P<i><sub>PA5530</sub>::lacZ</i> reporter. Cells were grown in LB media to an optical density at 600 nm (OD<sub>600</sub>) value of 0.3. They were then challenged with α-KG or no substrate and allowed to grow for an additional 60 min post-induction. Data points represent mean values±the standard deviations (<i>n</i>=3). Analysis of variance was performed by using Dunnett’s post-hoc test (α value of 0.05) to identify significant differences (<i>P</i>&lt;0.0001; marked with an asterisk).</div>
</figcaption>
</figure>
<figure class="figure-item "
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data-slide-index="1"
data-label-slug="fig-2">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/800bc5b8cfd9/mic-166-867-g002.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-1"
role="button"
data-image-width="734"
data-image-height="446"
data-image-alt="Fig. 2."
data-pmc-id="PMC7654738"
data-figure-id="F2">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/be0b69d3d67f/mic-166-867-g002.gif"
alt="Fig. 2." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="734">
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 2. </p>
</strong>
<div class="figure-caption-contents"><p> MifS activity is induced by… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 2. </p>
</strong>
<div class="figure-caption-contents"><p> MifS activity is induced by α-KG. <italic xmlns:xlink="http://www.w3.org/1999/xlink"> <ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093"> E. coli </ext-link> cells harbouring the P <i> <sub> PA5530… </sub> </i> </italic> </p></div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 2.
</strong>
<div class="figure-caption-contents">MifS activity is induced by α-KG. <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the P<i><sub>PA5530</sub>::lacZ</i> reporter and the <i>mifSR</i> genes were grown in LB media to an OD<sub>600</sub> value of 0.3 and induced with a panel of dicarboxylates at a final concentration of 20 mM. LacZ expression levels were measured 60 min post-induction. Data points represent mean values±the standard deviations (<i>n</i>=3). Analysis of variance was performed by using Dunnett’s post-hoc test (α value of 0.05) to identify significant differences (<i>P</i>&lt;0.0001; marked with an asterisk).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="2"
data-label-slug="fig-3">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/fe8ee81fbb55/mic-166-867-g003.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-2"
role="button"
data-image-width="787"
data-image-height="480"
data-image-alt="Fig. 3."
data-pmc-id="PMC7654738"
data-figure-id="F3">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/b693427f46ef/mic-166-867-g003.gif"
alt="Fig. 3." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="787">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="480">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 3. </p>
</strong>
<div class="figure-caption-contents"><p> MifS sensor domain has an… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 3. </p>
</strong>
<div class="figure-caption-contents"><p> MifS sensor domain has an α-KG binding pocket. (a) Structural model of the… </p></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 3.
</strong>
<div class="figure-caption-contents">MifS sensor domain has an α-KG binding pocket. (a) Structural model of the MifS periplasmic sensor domain showing the two mixed α/β fold PDC domains, each containing a central β-sheet surrounded by three α-helices. The α-KG molecule is shown in blue bound to the distal PDC domain. The homology model of the MifS sensor domain was constructed using the SWISS-MODEL homology server [17]. (b) Model of the MifS binding pocket showing α-KG interacting with binding pocket residues via predicted hydrogen bonds to the side-chains and backbones of the binding pocket residues. α-KG was docked into the MifS sensor domain using AutoDock Vina [18] and protein–ligand hydrogen bonds were predicted using PyMOL [19].</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="3"
data-label-slug="fig-4">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/3297dd8fc757/mic-166-867-g004.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="630"
data-image-height="445"
data-image-alt="Fig. 4."
data-pmc-id="PMC7654738"
data-figure-id="F4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/8bb479bc32ac/mic-166-867-g004.gif"
alt="Fig. 4." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="630">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="445">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 4. </p>
</strong>
<div class="figure-caption-contents"><p> The MifS sensor domain is… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 4. </p>
</strong>
<div class="figure-caption-contents"><p> The MifS sensor domain is required for response to α-KG. Mutations were introduced… </p></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 4.
</strong>
<div class="figure-caption-contents">The MifS sensor domain is required for response to α-KG. Mutations were introduced into the sensor domain of MifS. The wild-type <i>mifS</i> gene or one of the mutated <i>mifS</i> genes were introduced into <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the <i>mifR</i> gene and the P<i><sub>PA5530</sub>::lacZ</i> reporter. Cells were grown in LB media to an OD<sub>600</sub> value of 0.3 and challenged with α-KG (black) or no substrate (white). LacZ expression levels were measured 60 min post-induction. Data points represent mean values±the standard deviations (<i>n</i>=3). Analysis of variance was performed by using Dunnett’s post-hoc test (α value of 0.05) to identify significant differences (<i>P</i>&lt;0.0001; marked with an asterisk).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="4"
data-label-slug="fig-5">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/fe8044df3aae/mic-166-867-g005.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="689"
data-image-height="363"
data-image-alt="Fig. 5."
data-pmc-id="PMC7654738"
data-figure-id="F5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/46bda82e1ecf/mic-166-867-g005.gif"
alt="Fig. 5." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="689">
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 5. </p>
</strong>
<div class="figure-caption-contents"><p> MifS sensor domain residue Phe162… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 5. </p>
</strong>
<div class="figure-caption-contents"><p> MifS sensor domain residue Phe162 required for substrate specificity. (a) Alignment of substrate… </p></div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 5.
</strong>
<div class="figure-caption-contents">MifS sensor domain residue Phe162 required for substrate specificity. (a) Alignment of substrate binding pocket sequences of MifS, <i>Vc</i>_DctB and COG4191 (DctB family consensus sequence). Residues conserved in all three sequences are shown in boxes. Residues required for α-KG response in MifS are shown in bold. Residues required for succinate binding in <i>Vc</i>_DctB are indicated with a triangle. Alignment was generated using <sc>clustal</sc> Omega [56] and displayed using ESPript 3.0 [57]. (b) <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the wild-type <i>mifS</i> gene or the <i>mifSF162Y</i> gene along with the <i>mifR</i> gene and P<i><sub>PA5530</sub>::lacZ</i> reporter were grown in LB media to an OD<sub>600</sub> value of 0.3 and induced with a panel of dicarboxylates at a final concentration of 20 mM. (c) Wild-type MifS or a MifS variant with a mutation resembling the <i>Vc</i>_DctB binding pocket was expressed in <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the <i>mifR</i> gene and the P<i><sub>PA5530</sub>::lacZ</i> reporter. Cells were grown in LB media to an OD<sub>600</sub> value of 0.3 and challenged with succinate or no substrate. LacZ expression levels were measured 60 min post-induction. Data points represent mean values±the standard deviations (<i>n</i>=3). Analysis of variance was performed by using Dunnett’s post-hoc test (α value of 0.05) to identify significant differences (<i>P</i>&lt;0.0001; marked with an asterisk).</div>
</figcaption>
</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="5"
data-label-slug="fig-6">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/957817320cab/mic-166-867-g006.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="787"
data-image-height="370"
data-image-alt="Fig. 6."
data-pmc-id="PMC7654738"
data-figure-id="F6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81d7/7654738/6c46b26a9d90/mic-166-867-g006.gif"
alt="Fig. 6." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="787">
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> Fig. 6. </p>
</strong>
<div class="figure-caption-contents"><p> Linker domain involved in MifS… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 6. </p>
</strong>
<div class="figure-caption-contents"><p> Linker domain involved in MifS α-KG response. (a) Alignment of cytoplasmic linker domain… </p></div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 6.
</strong>
<div class="figure-caption-contents">Linker domain involved in MifS α-KG response. (a) Alignment of cytoplasmic linker domain sequences of MifS and COG4191 (DctB family consensus sequence). Positions with identical residues are boxed. Alignment was generated using <sc>clustal</sc> Omega [56] and displayed using ESPript 3.0 [57]. (b) Helical wheel representation of one heptad of the linker domain spanning from residue 336 to residue 342. The interactions between the a position L336 and the d position L339 of two adjacent helices are demonstrated by the helical wheel representation, generated using DrawCoil 1.0 (<ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://www.grigoryanlab.org/drawcoil">http://www.grigoryanlab.org/drawcoil</ext-link>). (c) Mutations were introduced into the linker domain of MifS. The wild-type <i>mifS</i> gene or one of the mutated <i>mifS</i> genes were introduced into <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.3093">E. coli</ext-link>
</i> cells harbouring the <i>mifR</i> gene and the P<i><sub>PA5530</sub>::lacZ</i> reporter. Cells were grown in LB media to an OD<sub>600</sub> value of 0.3 and challenged with α-KG or no substrate. LacZ expression levels were measured 60 min post-induction. Data points represent mean values±the standard deviations (<i>n</i>=3). Analysis of variance was performed by using Dunnett’s post-hoc test (α value of 0.05) to identify significant differences (<i>P</i>&lt;0.0001; marked with an asterisk).</div>
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aria-describedby="figure-caption-6"
role="button"
data-image-width="787"
data-image-height="523"
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data-pmc-id="PMC7654738"
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<strong class="figure-label">
<p> Fig. 7. </p>
</strong>
<div class="figure-caption-contents"><p> MifS shares sequence and structural… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig. 7. </p>
</strong>
<div class="figure-caption-contents"><p> MifS shares sequence and structural homology with DctB. A structural alignment of the… </p></div>
</div>
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig. 7.
</strong>
<div class="figure-caption-contents">MifS shares sequence and structural homology with DctB. A structural alignment of the MifS substrate binding pocket with that of <italic xmlns:xlink="http://www.w3.org/1999/xlink">
<ext-link ext-link-type="uri" xlink:href="http://doi.org/10.1601/nm.2947">V. cholerae</ext-link>
</i> DctB shows that the sensor domains of these two histidine kinases have several conserved residues. Structural alignment performed using PyMOL [19].</div>
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<h2 class="title">
Related information
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<ul class="related-links-list"><li><a title="Link to related Genes" href="https://www.ncbi.nlm.nih.gov/gene?linkname=pubmed_gene&amp;from_uid=32553056"
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Gene
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Gene (GeneRIF)
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MedGen
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