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<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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr">
<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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr">
<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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr">
<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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr">
<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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr">
<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="36759782">
<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="https://bmcmicrobiol.biomedcentral.com/articles/10.1186/s12866-023-02783-3"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3196&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=36759782&amp;db=pubmed&amp;nlmid=100966981"
title="See full text options at BioMed Central"
data-ga-category="full_text"
data-ga-action="BioMed Central"
data-ga-label="36759782"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--s3-service-broker-live-ddda94b7-dbb0-4917-917d-776dae91ebba.s3.amazonaws.com-bmc-linkout-fulltext.png" alt="BioMed Central full text link"><span class="text">
BioMed Central
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/36759782/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=36759782&amp;db=pubmed&amp;nlmid=100966981"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="36759782"
><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="/36759782/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/36759782/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="36759782" 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%2F36759782%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="bP0jWUQnYGHDhqxQ1siDeikWZr64eUR7sCKozKjJyhu7bRDGELVFIqQAxvOg7vpr"><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"
id="collections-action-dialog-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"
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SigB modulates expression of novel SigB regulon members via Bc1009 in non-stressed and heat-stressed cells revealing its alternative roles in Bacillus cereus
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SigB modulates expression of novel SigB regulon members via Bc1009 in non-stressed and heat-stressed cells revealing its alternative roles in Bacillus cereus
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<li data-affiliation-id="short-view-affiliation-4"
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><sup class="key">4</sup> Center for Synthetic Microbiology (SYNMIKRO) &amp; Department of Chemistry, Philipps-University Marburg, Karl-Von-Frisch-Strasse 14, 35043, Marburg, Germany.</li>
<li data-affiliation-id="short-view-affiliation-5"
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><sup class="key">5</sup> Food Microbiology, Wageningen University and Research, PO Box 8129, 6700 EV, Wageningen, The Netherlands. tjakko.abee@wur.nl.</li>
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Abstract
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Background:
</strong>
The Bacillus cereus Sigma B (SigB) dependent general stress response is activated via the two-component RsbKY system, which involves a phosphate transfer from RsbK to RsbY. It has been hypothesized that the Hpr-like phosphocarrier protein (Bc1009) encoded by bc1009 in the SigB gene cluster may play a role in this transfer, thereby acting as a regulator of SigB activation. Alternatively, Bc1009 may be involved in the activation of a subset of SigB regulon members.
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Results:
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We first investigated the potential role of bc1009 to act as a SigB regulator but ruled out this possibility as the deletion of bc1009 did not affect the expression of sigB and other SigB gene cluster members. The SigB-dependent functions of Bc1009 were further examined in B. cereus ATCC14579 via comparative proteome profiling (backed up by transcriptomics) of wt, Δbc1009 and ΔsigB deletion mutants under heat stress at 42 °C. This revealed 284 proteins displaying SigB-dependent alterations in protein expression levels in heat-stressed cells, including a subgroup of 138 proteins for which alterations were also Bc1009-dependent. Next to proteins with roles in stress defense, newly identified SigB and Bc1009-dependent proteins have roles in cell motility, signal transduction, transcription, cell wall biogenesis, and amino acid transport and metabolism. Analysis of lethal stress survival at 50 °C after pre-adaptation at 42 °C showed intermediate survival efficacy of Δbc1009 cells, highest survival of wt, and lowest survival of ΔsigB cells, respectively. Additional comparative proteome analysis of non-stressed wt and mutant cells at 30 °C revealed 96 proteins with SigB and Bc1009-dependent differences in levels: 51 were also identified under heat stress, and 45 showed significant differential expression at 30 °C. This includes proteins with roles in carbohydrate/ion transport and metabolism. Overlapping functions at 30 °C and 42 °C included proteins involved in motility, and ΔsigB and Δbc1009 cells showed reduced motility compared to wt cells in swimming assays at both temperatures.
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Conclusion:
</strong>
Our results extend the B. cereus SigB regulon to &gt; 300 members, with a novel role of SigB-dependent Bc1009 in the activation of a subregulon of &gt; 180 members, conceivably via interactions with other transcriptional regulatory networks.
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Keywords:
</strong>
Adaptive general stress response; Amino acid metabolisms; Metabolic crosstalk; Motility; Phosphocarrier protein; SigB baseline function; SigB subregulon; Sporulation; Virulence.
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<p> Fig. 1 </p>
</strong>
<div class="figure-caption-contents"><p> SigB gene cluster in <i> Bacillus… </i> </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
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<p> Fig. 1 </p>
</strong>
<div class="figure-caption-contents"><p> SigB gene cluster in <i> Bacillus cereus </i> and differential gene expression after heat shock.… </p></div>
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<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Fig. 1
</strong>
<div class="figure-caption-contents">SigB gene cluster in <i>Bacillus cereus</i> and differential gene expression after heat shock. <b>A-</b> SigB gene cluster arrangement in <i>B. cereu</i>s ATCC14579. <b>B-</b> Change in expression of genes belonging to the <i>sigB</i> gene cluster for wt and Δ<i>sigB</i> mutant after 20 min of heat shock at 42 °C compared to the non-heat-stressed condition at 30 °C. <i>N</i>=4, <i>p</i>&lt;0.001. <b>C-</b> Change in expression of genes belonging to the SigB gene cluster expression for wt and Δ<i>bc1009</i> mutant after heat shock at 42 °C compared to the non-heat-stressed condition at 30 °C; Black bar - wt; white bar - Δ<i>sigB</i> mutant; grey bar - Δ<i>bc1009</i> mutant. * indicates the absence of gene activity. <i>N</i>=4, <i>p</i>&lt;0.001, refers to the difference in expression values of wt vs. either Δ<i>sigB</i> or Δ<i>bc1009</i> mutant</div>
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<p> Fig. 2 </p>
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<div class="figure-caption-contents"><p> Proteomics analyses of <i> B. cereus… </i> </p></div>
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</div>
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<p> Fig. 2 </p>
</strong>
<div class="figure-caption-contents"><p> Proteomics analyses of <i> B. cereus </i> ATCC14579 wt, Δ <i> sigB, </i> and Δ <i> bc1009 </i> mutants… </p></div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Fig. 2
</strong>
<div class="figure-caption-contents">Proteomics analyses of <i>B. cereus</i> ATCC14579 wt, Δ<i>sigB,</i> and Δ<i>bc1009</i> mutants upon heat shock. Left: Venn diagram showing 429 significantly induced (upregulated) proteins in <i>B. cereus</i> heat-stressed wt cells (black circle) compared to the non-heat-stressed wt cells at 30 °C (see Supplementary Table S2A). White circle: 175 heat-induced proteins in wt that are SigB-dependent, i.e., proteins that show upregulation of&gt;0.6 log<sub>2</sub> fold change in wt/<i>ΔsigB</i> cells upon heat shock compared to the non-heat-stressed condition at 30 °C. Grey circle: For 98 proteins, the heat-mediated increase in level in the wt is dependent on SigB and Bc1009, i.e., proteins that show upregulation of&gt;0.6 log<sub>2</sub> fold change in wt/Δ<i>sigB</i> and wt/Δ<i>bc1009</i> cells upon heat shock compared to the non-heat-stressed condition at 30 °C (see Table 2 for the complete list of proteins). Right: Venn diagram showing 435 significantly downregulated proteins in <i>B. cereus</i> heat-stressed wt cells (black circle) compared to the non-heat-stressed wt cells at 30 °C (Supplementary Table S2A). White circle: 109 downregulated proteins in heat-stressed wt cells vs. non-heat-stressed wt cells are SigB-dependent, i.e., proteins that show downregulation of&gt;0.6 log<sub>2</sub> fold change in wt/Δ<i>sigB</i> cells upon heat shock compared to the non-heat-stressed condition at 30 °C. Grey circle: For 40 proteins, reduction in level in wt cells upon heat shock was dependent on SigB and Bc1009, i.e., proteins that show downregulation of&gt;0.6 log<sub>2</sub> fold change in wt/Δ<i>sigB</i> and wt/Δ<i>bc1009</i> cells upon heat shock vs. non-heat-stressed condition (see Table 3). The underlying transcriptome data supporting this figure are presented in Supplementary Table S2B</div>
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<img class="figure-thumb" itemprop="thumbnail"
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<p> Fig. 3 </p>
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<div class="figure-caption-contents"><p> SigB and SigB (and Bc1009)-dependent… </p></div>
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<p> Fig. 3 </p>
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<div class="figure-caption-contents"><p> SigB and SigB (and Bc1009)-dependent induced and downregulated proteins in wt cells after… </p></div>
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Fig. 3
</strong>
<div class="figure-caption-contents">SigB and SigB (and Bc1009)-dependent induced and downregulated proteins in wt cells after heat shock (30 °C to 42 °C) versus before heat shock and their cluster of orthologous group (COG). <b>A</b>- log<sub>2</sub> fold change in protein expression. Positive and negative fold change values indicate induced and downregulated proteins, respectively. Numbers indicate the top 30 induced (Table 2) or top 20 downregulated (Table 3) proteins, respectively. The <i>p</i>-value threshold for each protein was&lt;0.05, <i>N</i>=4. Grey symbols: induced/downregulated proteins that are SigB (and Bc1009-dependent; i.e., proteins that show up(or down) regulation of&gt;0.6 log<sub>2</sub> fold change in wt/Δ<i>sigB</i> and wt/Δ<i>bc1009</i> cells upon heat shock vs. non-heat-stressed condition. White symbols: induced/downregulated proteins that are SigB-dependent; i.e., proteins that show up(or down) regulation of&gt;0.6 log<sub>2</sub> fold change in wt/Δ<i>sigB</i> cells upon heat shock vs. non-heat-stressed condition. <b>B</b>- cluster of orthologous group (COG) function for both SigB-dependent induced (Right) and downregulated proteins (Left) in wt cells upon heat shock. The number on the x-axis indicates the total number of induced/downregulated proteins, and the negative sign indicates downregulation. Grey bar: induced/downregulated proteins after heat shock that are SigB (and Bc1009)-dependent, i.e., proteins that show up(or down) regulation of&gt;0.6 log<sub>2</sub> fold change in wt/<i>ΔsigB</i> and wt/Δ<i>bc1009</i> cells; White bar: induced/downregulated proteins that are SigB-dependent, i.e., proteins that show up(or down) regulation of&gt;0.6 log<sub>2</sub> fold change only in wt/Δ<i>sigB</i> cells respectively, compared to the expression in the non-heat-stressed condition at 30 °C. The underlying transcriptome data supporting this figure are presented in Supplementary Table S3B</div>
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<strong class="figure-label">
<p> Fig. 4 </p>
</strong>
<div class="figure-caption-contents"><p> The relative survival of <i> B.… </i> </p></div>
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<strong class="figure-label">
<p> Fig. 4 </p>
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<div class="figure-caption-contents"><p> The relative survival of <i> B. cereus </i> wt, Δ <i> sigB, </i> and Δ <i> bc1009 </i> mutants… </p></div>
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Fig. 4
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<div class="figure-caption-contents">The relative survival of <i>B. cereus</i> wt, Δ<i>sigB,</i> and Δ<i>bc1009</i> mutants upon lethal heat exposure at 50°C. The relative survival at 50 °C of heat-preadapted cells (30 °C to 42 °C for 45 min) of <i>B. cereus</i> wt (filled circle), Δ<i>sigB</i> (filled triangle), and Δ<i>bc1009</i> cells (filled square) compared to cells that were not preadapted to heat (42 °C) (wt- open circle; Δ<i>sigB</i>- open triangle; Δ<i>bc1009</i>- open square) for 120 min. <i>N</i>=4. <i>p</i>&lt;0.001 for time point at 100 min and 120 min when comparing wt and the two mutant strains. Error bars show the standard deviation of four biological replicates</div>
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<strong class="figure-label">
<p> Fig. 5 </p>
</strong>
<div class="figure-caption-contents"><p> SigB-dependent induced and downregulated proteins… </p></div>
</div>
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<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> SigB-dependent induced and downregulated proteins for Δ <i> sigB </i> and Δ <i> bc1009 </i> mutants compared… </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">SigB-dependent induced and downregulated proteins for Δ<i>sigB</i> and Δ<i>bc1009</i> mutants compared to wt cells at 30°C and their cluster of orthologous group (COG). <b>A</b>- log<sub>2</sub> fold change in protein expression. Positive and negative fold change values indicate induced and downregulated proteins, respectively. Grey symbols: induced/downregulated proteins that are SigB (and Bc1009)- dependent, i.e., differentially expressed proteins in Δ<i>sigB</i> and Δ<i>bc1009</i> mutants compared to wt cells at 30 °C; the numbers indicate the top 30 induced proteins (positive x-axis) or top 20 downregulated proteins (negative x-axis), with details listed in Table 4. White symbols: induced/downregulated proteins at 30 °C that are either only SigB-dependent or Bc1009-dependent, i.e., differentially expressed proteins in Δ<i>sigB</i> or Δ<i>bc1009</i> mutants compared to wt cells at 30 °C (see details in Supplementary Table S5A). The threshold of the <i>p</i>-value for each protein was&lt;0.05. <i>N</i>=4. <b>B</b>- cluster of orthologous group (COG) function for SigB (and Bc1009)-induced and downregulated proteins. The number on the x-axis indicates the number of induced/downregulated proteins in respective COG groups. The underlying transcriptome data supporting this figure are presented in Supplementary Table S5B</div>
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<p> Fig. 6 </p>
</strong>
<div class="figure-caption-contents"><p> Motility phenotype of wt, Δ… </p></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
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<p> Fig. 6 </p>
</strong>
<div class="figure-caption-contents"><p> Motility phenotype of wt, Δ <i> sigB, </i> and Δ <i> bc1009 </i> mutants. <b> A </b> - the… </p></div>
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<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Fig. 6
</strong>
<div class="figure-caption-contents">Motility phenotype of wt, Δ<i>sigB,</i> and Δ<i>bc1009</i> mutants. <b>A</b>- the motility of wt, Δ<i>sigB,</i> and Δ<i>bc1009</i> was compared on Brain Heart Infusion (BHI) agar with 0.25% agar and indicated by the colony diameter (mm) formed on the agar after 24 h incubation. Black bar: wt; Grey bar: Δ<i>sigB</i> mutant; white bar: Δ<i>bc1009</i> mutant; light grey bar: Δ<i>flgG</i> mutant (negative control without flagella). The motility of all cells was tested under three different conditions, 1) at 30 °C for 24 h; 2) upon heat shock from 30 °C to 42 °C for 30 min, and back to 30 °C for 24 h; and 3) at 42 °C for 24 h. The dotted line shows the maximum plate size. <b>B</b>- Colony of wt, Δ<i>sigB,</i> Δ<i>bc1009,</i> and Δ<i>flgG</i> cells on 0.25% BHI agar at 30 °C after 24 h. Error bars indicates the standard deviation of colony diameters of four biological replicates</div>
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<p> Fig. 7 </p>
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<div class="figure-caption-contents"><p> Activation model of <i> Bacillus cereus… </i> </p></div>
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<p> Fig. 7 </p>
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<div class="figure-caption-contents"><p> Activation model of <i> Bacillus cereus </i> SigB and the SigB-dependent Bc1009 subregulon. The RsbKY… </p></div>
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<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Fig. 7
</strong>
<div class="figure-caption-contents">Activation model of <i>Bacillus cereus</i> SigB and the SigB-dependent Bc1009 subregulon. The RsbKY two-component system mediates the SigB general stress response in <i>B. cereus.</i> Under heat shock (temperature shift from 30 °C to 42 °C), SigB is induced (bold arrow) via the RsbKY signaling cascade and directly regulates the Hpr-like phosphocarrier protein Bc1009 to mediate amino acid transport and metabolism, cell motility, signal transduction, cell wall/membrane/envelope biogenesis and transcription. At 30 °C, the baseline level of SigB also appears to regulate Bc1009 (non-bold arrow) to mediate similar functions as described for the heat shock condition, except for amino acid transport and metabolism (which switch to carbohydrate/ion transport and metabolism at 30 °C instead, indicated in dark grey and light grey box, respectively). Upon SigB activation, other SigB regulon members not controlled by Bc1009 are also activated (see main text, Tables 2 and 3, and Supplementary Tables S5A and B). White color circles: known SigB signalling pathway as described in the literature [5, 12, 15, 16], Grey and Black circles and boxes: new insight obtained in the current study (see text for details)</div>
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Lund T, De Buyser ML, Granum PE. A new cytotoxin from Bacillus cereus that may cause necrotic enteritis. Mol Microbiol. 2000;38:254261. doi: 10.1046/j.1365-2958.2000.02147.x.
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Ceuppens S, Rajkovic A, Hamelink S, Van de Wiele T, Boon N, Uyttendaele M. Enterotoxin production by Bacillus cereus under gastrointestinal conditions and their immunological detection by commercially available kits. Foodborne Pathog Dis. 2012;9:11301136. doi: 10.1089/fpd.2012.1230.
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Desriac N, Broussolle V, Postollec F, Mathot A-G, Sohier D, Coroller L, et al. Bacillus cereus cell response upon exposure to acid environment: toward the identification of potential biomarkers. Front Microbiol. 2013;4:284. doi: 10.3389/fmicb.2013.00284.
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