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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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z">
<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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z">
<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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z">
<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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z">
<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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z">
<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="39343004">
<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://linkinghub.elsevier.com/retrieve/pii/S0021-9258(24)02322-6"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=3048&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=39343004&amp;db=pubmed&amp;nlmid=2985121R"
title="See full text options at Elsevier Science"
data-ga-category="full_text"
data-ga-action="Elsevier Science"
data-ga-label="39343004"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--linkinghub.elsevier.com-ihub-images-elsevieroa.png" alt="Elsevier Science full text link"><span class="text">
Elsevier Science
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/39343004/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=39343004&amp;db=pubmed&amp;nlmid=2985121R"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="39343004"
><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="/39343004/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/39343004/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="39343004" 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%2F39343004%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="Vr5XoJVdwSuyUZnB5ST42nz46eMPsVtfceP21zoz6th2OqtrIbw6wv5IEiu1lw1z"><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"
data-ga-category="collections_button"
data-ga-action="create_collection"
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Nonfunctional coq10 mutants maintain the ERMES complex and reveal true phenotypes associated with the loss of the coenzyme Q chaperone protein Coq10
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Abstract
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<p>
Coenzyme Q (CoQ) is a redox-active lipid molecule that acts as an electron carrier in the mitochondrial electron transport chain. In Saccharomyces cerevisiae, CoQ is synthesized in the mitochondrial matrix by a multisubunit protein-lipid complex termed the CoQ synthome, the spatial positioning of which is coordinated by the endoplasmic reticulum-mitochondria encounter structure (ERMES). The MDM12 gene encoding the cytosolic subunit of ERMES is coexpressed with COQ10, which encodes the putative CoQ chaperone Coq10, via a shared bidirectional promoter. Deletion of COQ10 results in respiratory deficiency, impaired CoQ biosynthesis, and reduced spatial coordination between ERMES and the CoQ synthome. While Coq10 protein content is maintained upon deletion of MDM12, we show that deletion of COQ10 by replacement with a HIS3 marker results in diminished Mdm12 protein content. Since deletion of individual ERMES subunits prevents ERMES formation, we asked whether some or all of the phenotypes associated with COQ10 deletion result from ERMES dysfunction. To identify the phenotypes resulting solely due to the loss of Coq10, we constructed strains expressing a functionally impaired (coq10-L96S) or truncated (coq10-R147) Coq10 isoform using CRISPR-Cas9. We show that both coq10 mutants preserve Mdm12 protein content and exhibit impaired respiratory capacity like the coq10Δ mutant, indicating that Coq10's function is vital for respiration regardless of ERMES integrity. Moreover, the maintenance of CoQ synthome stability and efficient CoQ biosynthesis observed for the coq10-R147 mutant suggests these deleterious phenotypes in the coq10Δ mutant result from ERMES disruption. Overall, this study clarifies the role of Coq10 in modulating CoQ biosynthesis.
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Keywords:
</strong>
CoQ synthome; Coq10; Mdm12; START domain; Saccharomyces cerevisiae; coenzyme Q; endoplasmic reticulum-mitochondria encounter structure; lipid; mitochondrial metabolism; ubiquinone.
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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">Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.</p>
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Figures
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<div class="figure-caption-contents"><b> CoQ biosynthesis in yeast requires… </b></div>
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<div class="figure-caption-contents"><b> CoQ biosynthesis in yeast requires the CoQ synthome to assemble adjacent to ERMES… </b></div>
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<div class="figure-caption-contents"><b>CoQ biosynthesis in yeast requires the CoQ synthome to assemble adjacent to ERMES contact sites.</b><i>A</i>, the proposed CoQ biosynthetic pathway in <i>Saccharomyces cerevisiae</i>. <i>B</i>, schematic depicting the CoQ synthome positioned adjacent to the ERMES complex. CoQ synthome members are represented in purple and ERMES components are highlighted in <i>turquoise</i>. Coq1, Coq2, and Coq10 (<i>gray</i>) are not members of the CoQ synthome but are still required to observe efficient CoQ<sub>6</sub> biosynthesis. This image was generated using <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://BioRender.com">BioRender.com</ext-link>. CoQ, coenzyme Q; ERMES, endoplasmic reticulum-mitochondria encounter structure.</div>
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<div class="figure-caption-contents"><b> The Mdm12 and Mmm1 polypeptides… </b></div>
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<p> Figure 2 </p>
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<div class="figure-caption-contents"><b> The Mdm12 and Mmm1 polypeptides are depleted in yeast <i> coq10 </i> Δ mutants. </b> <i> A… </i></div>
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<div class="figure-caption-contents"><b>The Mdm12 and Mmm1 polypeptides are depleted in yeast <i>coq10</i>Δ mutants.</b><i>A</i>, aliquots of crude mitochondria (25 μg) from WT, <i>coq10</i>Δ, <i>coq11</i>Δ, and <i>coq10</i>Δ<i>coq11</i>Δ yeast strains were subjected to 10% Tris-glycine SDS-PAGE. Immunoblotting was performed with antisera against the indicated ERMES subunits (Mmm1, Mdm10, and Mdm12), and yeast harboring the corresponding deletions were used as negative controls (<i>ERMES</i>Δ). Malate dehydrogenase (Mdh1) was used as a loading control. Data are representative of three biological replicates. <i>B</i><i>D</i>, ImageJ was used to quantify triplicate band intensities of select ERMES subunits. Band intensities were normalized to Mdh1 and plotted as percentage of the WT control. The data depict the mean ± SD of three biological replicates. The statistical significance compared with WT is represented by the <i>red asterisks</i>; , <i>p</i> &lt; 0.05 and , <i>p</i> &lt; 0.0001. ERMES, endoplasmic reticulum-mitochondria encounter structure.</div>
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<div class="figure-caption-contents"><b> Structural prediction of <i> Saccharomyces cerevisiae… </i> </b></div>
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<div class="figure-caption-contents"><b> Structural prediction of <i> Saccharomyces cerevisiae </i> Coq10 and multiple sequence alignment with human COQ10… </b></div>
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<div class="figure-caption-contents"><b>Structural prediction of <i>Saccharomyces cerevisiae</i> Coq10 and multiple sequence alignment with human COQ10 orthologs highlight residues targeted for mutagenesis.</b><i>A</i>, multiple sequence alignment of <i>S. cerevisiae</i> Coq10 (residues 31207) with the Coq10-L96S and Coq10-R147 mutant polypeptides constructed in this study and the human homologs COQ10A (residues 73244)/COQ10B (residues 64235). The yeast Coq10 polypeptide and orthologous human sequences were obtained from Universal Protein Knowledgebase (UniProtKB). The multiple sequence alignment was constructed using the ClustalW package of Clustal Omega (59) and visualized in JalView2 (60). Conservation of each residue is indicated by degree of shading, which represents 80%, 60% and 40% percent sequence identity from <i>darkest to lightest shade</i>, respectively. Residues targeted for mutagenesis in this study are indicated with an <i>inverted triangle</i>, and <i>asterisks</i> indicate residues deemed critical for ligand binding in previous studies (11). <i>B</i>, location of the Coq10 residues targeted for mutagenesis by CRISPR-Cas9 (shown in <i>green</i>) within the context of the <i>COQ10</i> ORF (<i>top</i>) and the AlphaFold predicted structure for <i>S. cerevisiae</i> Coq10 (<i>bottom</i>, AF-Q08058-F1). The region shown in <i>gray</i> represents the truncation that results from introducing the <i>coq10-R147/N149</i> double mutation. <i>C</i>, schematic depicting the head-to-head positioning of <i>COQ10</i> (<i>purple</i>) and <i>MDM12</i> (<i>gray</i>) within the context of <i>S. cerevisiae</i> chromosome XV (<i>green</i>). Notably, these two genes are separated by only 173 bps, suggesting deletion of one gene could impact the expression of the other, and <i>vice versa</i>.</div>
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<div class="figure-caption-contents"><b> Mdm12 and Mmm1 protein content… </b></div>
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<div class="figure-caption-contents"><b> Mdm12 and Mmm1 protein content is preserved in strains expressing either the Coq10-L96S… </b></div>
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<div class="figure-caption-contents"><b>Mdm12 and Mmm1 protein content is preserved in strains expressing either the Coq10-L96S or Coq10-R147 mutant polypeptide.</b><i>A</i>, aliquots of crude mitochondria (25 μg) from the indicated yeast strains were subjected to 10% Tris-glycine SDS-PAGE. Immunoblotting was performed with antisera against the indicated ERMES subunits (Mmm1, Mdm10, and Mdm12), and mitochondria from yeast harboring the corresponding deletions were used as negative controls (<i>ERMES</i>Δ). Malate dehydrogenase (Mdh1) was used as a loading control. Data are representative of three biological replicates. <i>B</i><i>D</i>, ImageJ was used to quantify triplicate band intensities of the indicated ERMES proteins. Band intensities were normalized to Mdh1 and plotted as percentage of the WT control. The data depict mean ± SD of three biological replicates, and the statistical significance compared with WT is represented by the <i>red asterisks</i> , <i>p</i> &lt; 0.05 in panel <i>B</i>, or by the stated <i>p</i> value in <i>D</i>. <i>E</i>, 12.5 μg of crude mitochondria were separated on 12% Tris-glycine SDS-PAGE and immunoblotting was performed using Coq10 antisera. An aliquot of mitochondria from the <i>coq10</i>Δ yeast was used as a negative control. <i>F</i>, ImageJ was used to quantify triplicate band intensities for the Coq10 polypeptide. Band intensities were normalized to Mdh1 and plotted as percentage of the WT control. The data depict the mean ± SD of three biological replicates. The statistical significance compared with WT is represented by the <i>red asterisks</i>; , <i>p</i> &lt; 0.0001. ERMES, endoplasmic reticulum-mitochondria encounter structure.</div>
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<div class="figure-caption-contents"><b> Insertion of the <i> HIS3 </i> marker… </b></div>
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<div class="figure-caption-contents"><b> Insertion of the <i> HIS3 </i> marker at the <i> COQ10 </i> locus causes aberrant transcription into… </b></div>
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<div class="figure-caption-contents"><b>Insertion of the <i>HIS3</i> marker at the <i>COQ10</i> locus causes aberrant transcription into the neighboring <i>MDM12</i> gene.</b> The <i>bar graphs</i> show normalized read counts for <i>A</i>, <i>COQ10</i> and <i>B</i>, <i>MDM12</i> mRNAs based on reads obtained with Oxford Nanopore sequencing using a complementary DNA approach for three replicates per strain (see Experimental procedures). DESeq2 was used to quantify changes in gene expression of the different strains and for normalizing counts to library size. The p-adjusted is used to indicate significance. The data depict the mean ± SD of three biological replicates, and the statistical significance of <i>COQ10</i> read counts compared with WT is represented by the <i>red asterisks</i>, , <i>p</i> &lt; 0.01; and , <i>p</i> &lt; 0.0001. No significant differences were observed for the <i>MDM12</i> read counts. <i>C</i>, nanopore sequencing reads detected in the <i>COQ10</i> and <i>MDM12</i> regions for the indicated strains. Each sequencing read is represented by a horizontal gray line. No reads were detected for the <i>COQ10</i> ORF in the <i>coq10</i>Δ mutant because of the replacement of the <i>COQ10</i> ORF by the <i>HIS3</i> marker. Since the reads were aligned to the WT genome, reads corresponding to the <i>HIS3</i> gene are not shown. The <i>HIS3</i> marker is oriented in a way that positions the <i>HIS3</i> promoter adjacent to the 3 UTR of the deleted <i>COQ10</i> ORF.</div>
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<div class="figure-caption-contents"><b> The <i> coq10-L96S </i> and <i> coq10-R147 </i> mutants… </b></div>
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<div class="figure-caption-contents"><b> The <i> coq10-L96S </i> and <i> coq10-R147 </i> mutants display impaired respiratory growth similar to the <i> coq10… </i> </b></div>
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<div class="figure-caption-contents"><b>The <i>coq10-L96S</i> and <i>coq10-R147</i> mutants display impaired respiratory growth similar to the <i>coq10</i>Δ mutant that can be restored by deletion of <i>COQ11</i>.</b> Overnight cultures of the indicated yeast strains were diluted to an <i>A</i><sub>600</sub> = 0.2, and 2 μl of 5-fold serial dilutions were spotted onto fermentable (YPDextrose, YPD) or respiratory (YPGlycerol, YPG) medium. Plates were incubated at 30 °C for 2 or 3 days prior to imaging. Data are representative of three biological replicates.</div>
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<div class="figure-caption-contents"><b> The <i> coq10-R147 </i> mutant retains the… </b></div>
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<div class="figure-caption-contents"><b> The <i> coq10-R147 </i> mutant retains the ability to efficiently synthesize CoQ </b> <sub> <b> 6 </b> </sub> <b> . </b> Triplicates…</div>
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Figure 7
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<div class="figure-caption-contents"><b>The <i>coq10-R147</i> mutant retains the ability to efficiently synthesize CoQ</b><sub><b>6</b></sub><b>.</b> Triplicates of yeast cultured in 25 ml YPGal were labeled at an <i>A</i><sub>600</sub> 0.6 with 8 μg/ml <sup>13</sup>C<sub>6</sub>-pABA or ethanol as a vehicle control. Fifteen milliliters of each culture were harvested after 5 h, lipid extracted, and analyzed by LC-MS/MS. <i>A</i>, unlabeled <sup>12</sup>C-CoQ<sub>6</sub>; <i>B</i>, labeled <sup>13</sup>C<sub>6</sub>-CoQ<sub>6</sub>; <i>C</i>, total amount of CoQ<sub>6</sub> determined from the sum of <sup>12</sup>C-CoQ<sub>6</sub> (<i>white</i>) and <sup>13</sup>C<sub>6</sub>-CoQ<sub>6</sub> (<i>red</i>). The data depict the mean ± SD. The statistical significance as compared with the <i>coq10</i>Δ mutant is represented by the <i>black asterisks</i>; , <i>p</i> &lt; 0.05; , <i>p</i> &lt; 0.001; and , <i>p</i> &lt; 0.0001. CoQ, coenzyme Q; pABA, para-aminobenzoic acid; YPGal, yeast extract-peptone-galactose.</div>
</figcaption>
</figure>
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data-label-slug="figure-8">
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itemprop="contentUrl"
aria-describedby="figure-caption-7"
role="button"
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data-image-height="418"
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data-pmc-id="PMC11541779"
data-figure-id="fig8">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-7"
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</a>
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<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 8 </p>
</strong>
<div class="figure-caption-contents"><b> The <i> coq10-L96S </i> mutant has impaired… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 8 </p>
</strong>
<div class="figure-caption-contents"><b> The <i> coq10-L96S </i> mutant has impaired CoQ </b> <sub> <b> 6 </b> </sub> <b> biosynthetic efficiency in comparison to <i> coq10… </i> </b></div>
</div>
<figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 8
</strong>
<div class="figure-caption-contents"><b>The <i>coq10-L96S</i> mutant has impaired CoQ</b><sub><b>6</b></sub><b>biosynthetic efficiency in comparison to <i>coq10</i>Δ.</b> Triplicate of 25 ml cultures in YPGal were labeled at an <i>A</i><sub>600</sub> 0.6 with 8 μg/ml <sup>13</sup>C<sub>6</sub>-pABA or ethanol. 15 ml of each culture were harvested after 5 h, lipid extracted, and analyzed by LC-MS/MS. <i>A</i>, <sup>12</sup>C-HAB (<i>white</i>) and <sup>13</sup>C<sub>6</sub>-HAB (<i>red</i>) and <i>B</i>, <sup>12</sup>C-DMQ<sub>6</sub> (<i>white</i>) and <sup>13</sup>C<sub>6</sub>-DMQ<sub>6</sub> (<i>red</i>) were measured from whole-cell lipid extracts of the indicated yeast strains. Total HAB and DMQ<sub>6</sub> were determined from the sum of the respective labeled and unlabeled analytes. The data show mean ± SD. The statistical significance as compared to the <i>coq10</i>Δ mutant is represented by the <i>black asterisks</i>; , <i>p</i> &lt; 0.05; , <i>p</i> &lt; 0.01; , <i>p</i> &lt; 0.001; and , <i>p</i> &lt; 0.0001. CoQ, coenzyme Q; DMQ, demethoxy-Q; HAB, hexaprenylaminobenzoic acid; pABA, para-aminobenzoic acid; YPGal, yeast extract-peptone-galactose.</div>
</figcaption>
</figure>
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data-image-height="418"
data-image-alt="Figure 9"
data-pmc-id="PMC11541779"
data-figure-id="fig9">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-8"
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</a>
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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> Figure 9 </p>
</strong>
<div class="figure-caption-contents"><b> Decreased abundance of several Coq… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 9 </p>
</strong>
<div class="figure-caption-contents"><b> Decreased abundance of several Coq polypeptides in the <i> coq10-L96S </i> and <i> coq10-R147 </i> mutants can… </b></div>
</div>
<figcaption id="figure-caption-8" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 9
</strong>
<div class="figure-caption-contents"><b>Decreased abundance of several Coq polypeptides in the <i>coq10-L96S</i> and <i>coq10-R147</i> mutants can be rescued by deletion of <i>COQ11</i>.</b><i>A</i>, aliquots of crude mitochondria (12.5 μg) from the indicated yeast strains were subjected to 10% or 12% Tris-glycine SDS-PAGE. Crude mitochondria from <i>coq3</i>Δ<i>-coq9</i>Δ mutants were included as negative controls for Western blotting using antisera against each of the Coq polypeptides. Mitochondrial malate dehydrogenase (Mdh1) was included as a loading control. Data are representative of three biological replicates. <i>B</i>, ImageJ was used to quantify triplicate band intensities for each of the Coq polypeptides. Band intensities were normalized to Mdh1 and plotted as percentage of the WT control. The data depict the mean ± SD of three biological replicates. The statistical significance as compared with WT (<i>red asterisks</i>) or the <i>coq10</i>Δ mutant (<i>black asterisks</i>) are represented by, , <i>p</i> &lt; 0.05; , <i>p</i> &lt; 0.01; , <i>p</i> &lt; 0.001; and , <i>p</i> &lt; 0.0001. <i>C</i>, blots were quantified as in B. The data depict the mean ± SD of three biological replicates, and the statistical significance as compared with WT (<i>red asterisks</i>) or the <i>coq10</i>Δ<i>coq11</i>Δ mutant (<i>black asterisks</i>) are represented by ns, no significance; and , <i>p</i> &lt; 0.05.</div>
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data-label-slug="figure-10">
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data-figure-id="fig10">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-9"
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<div class="caption-wrap">
<strong class="figure-label">
<p> Figure 10 </p>
</strong>
<div class="figure-caption-contents"><b> Relative Coq11 protein abundance is… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 10 </p>
</strong>
<div class="figure-caption-contents"><b> Relative Coq11 protein abundance is significantly increased in strains harboring <i> coq10 </i> mutations. </b> <i> A… </i></div>
</div>
<figcaption id="figure-caption-9" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 10
</strong>
<div class="figure-caption-contents"><b>Relative Coq11 protein abundance is significantly increased in strains harboring <i>coq10</i> mutations.</b><i>A</i>, aliquots of crude mitochondria (12.5 μg) from the indicated yeast strains were subjected to 10% Tris-glycine SDS-PAGE. An aliquot of crude mitochondria from the <i>coq11</i>Δ mutant was included as a negative control for Western blotting against the Coq11 polypeptide. Mitochondrial malate dehydrogenase (Mdh1) was included as a loading control. The Mdh1 blot is replicated from Fig. 9<i>A</i> as the same sets of samples were used across all biological replicates when quantifying protein content for the individual Coq polypeptides. Data are representative of three biological replicates. <i>B</i>, ImageJ was used to quantify triplicate band intensities corresponding to Coq11. Band intensities were normalized to Mdh1 and plotted as percentage of the WT control. The data depict the mean ± SD of three biological replicates. The statistical significance compared with WT is represented by the <i>red asterisks</i>; , <i>p</i> &lt; 0.05; , <i>p</i> &lt; 0.01; , <i>p</i> &lt; 0.001.</div>
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<strong class="figure-label">
<p> Figure 11 </p>
</strong>
<div class="figure-caption-contents"><b> The <i> coq10-R147 </i> mutant maintains a… </b></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 11 </p>
</strong>
<div class="figure-caption-contents"><b> The <i> coq10-R147 </i> mutant maintains a stable CoQ synthome similar to the WT control.… </b></div>
</div>
<figcaption id="figure-caption-10" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Figure 11
</strong>
<div class="figure-caption-contents"><b>The <i>coq10-R147</i> mutant maintains a stable CoQ synthome similar to the WT control.</b> Aliquots (75 μg) of crude mitochondria isolated from WT, <i>coq10</i>Δ, <i>coq11</i>Δ, <i>coq10</i>Δ<i>coq11</i>Δ, <i>coq10-L96S</i>, <i>coq10-L96S coq11</i>Δ, <i>coq10-R147</i>, and <i>coq10-R147 coq11</i>Δ yeast were solubilized with digitonin and separated by 2D BN/SDS-PAGE. Proteins were transferred to polyvinylidene fluoride membranes, and the CoQ synthome was visualized using antisera against Coq9. Aliquots (25 μg) of intact crude mitochondria from each strain (M) and <i>coq9</i>Δ (<i>9</i>Δ) yeast were included as a loading control and negative control, respectively. BN, blue native; CoQ, coenzyme Q.</div>
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Okada K., Suzuki K., Kamiya Y., Zhu X., Fujisaki S., Nishimura Y., et al. Polyprenyl diphosphate synthase essentially defines the length of the side chain of ubiquinone. Biochim. Biophys. Acta. 1996;1302:217223.
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Frei B., Kim M.C., Ames B.N. Ubiquinol-10 is an effective lipid-soluble antioxidant at physiological concentrations. Proc. Natl. Acad. Sci. U. S. A. 1990;87:48794883.
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Awad A.M., Bradley M.C., Fernandez-Del-Rio L., Nag A., Tsui H.S., Clarke C.F. Coenzyme Q10 deficiencies: pathways in yeast and humans. Essays Biochem. 2018;62:361376.
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