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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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA">
<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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA">
<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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA">
<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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA">
<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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA">
<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="17993620">
<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://academic.oup.com/plcell/article-lookup/doi/10.1105/tpc.107.055319"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=7898&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=17993620&amp;db=pubmed&amp;nlmid=9208688"
title="See full text options at Silverchair Information Systems"
data-ga-category="full_text"
data-ga-action="Silverchair Information Systems"
data-ga-label="17993620"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--academic.oup.com-images-oup_pubmed.png" alt="Silverchair Information Systems full text link"><span class="text">
Silverchair Information Systems
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/17993620/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=17993620&amp;db=pubmed&amp;nlmid=9208688"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="17993620"
><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="/17993620/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/17993620/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="17993620" 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%2F17993620%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="QvVOhLtT1xPybG1dfUlsh2a2i87uqkpsF3V1VbZbGzukwKFj6hLpt0FePCcQxyVA"><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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The eukaryotic defense response posttranscriptional gene silencing (PTGS) is directed by short-interfering RNAs and thwarts invading nucleic acids via the RNA slicing activity of conserved ARGONAUTE (AGO) proteins. PTGS can be counteracted by exogenous or endogenous suppressors, including the cytoplasmic exoribonuclease XRN4, which also degrades microRNA (miRNA)-guided mRNA cleavage products but does not play an obvious role in development. Here, we show that the nuclear exoribonucleases XRN2 and XRN3 are endogenous PTGS suppressors. We also identify excised MIRNA loops as templates for XRN2 and XRN3 and show that XRN3 is critical for proper development. Independently, we identified the nucleotidase/phosphatase FIERY1 (FRY1) as an endogenous PTGS suppressor through a suppressor screen in a hypomorphic ago1 genetic background. FRY1 is one of six Arabidopsis thaliana orthologs of yeast Hal2. Yeast hal2 mutants overaccumulate 3'-phosphoadenosine 5'-phosphate, which suppresses the 5'--&gt;3' exoribonucleases Xrn1 and Rat1. fry1 mutant plants recapitulate developmental and molecular characteristics of xrn mutants and likely restore PTGS in ago1 hypomorphic mutants by corepressing XRN2, XRN3, and XRN4, thus increasing RNA silencing triggers. We anticipate that screens incorporating partially compromised silencing components will uncover additional PTGS suppressors that may not be revealed using robust silencing systems.
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data-pmc-id="PMC2174888"
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
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<strong class="figure-label">
<p> Figure 1. </p>
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<div class="figure-caption-contents"><p> Structure and Conservation of FRY1.… </p></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 1. </p>
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<div class="figure-caption-contents"><p> Structure and Conservation of FRY1. <b> (A) </b> <i> FRY1 </i> (At5g63980) gene structure. Exons are represented… </p></div>
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Figure 1.
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<div class="figure-caption-contents">Structure and Conservation of FRY1. <b>(A)</b> <i>FRY1</i> (At5g63980) gene structure. Exons are represented by black boxes. The positions of the ATG start and TGA stop codons are indicated. The location of <i>fry1</i> T-DNA insertions (triangles), nonsense mutations, and missense mutations with the corresponding amino acid changes are indicated above or below the gene structure. <b>(B)</b> Phylogeny of the FRY1 protein. In black are plants (At, <i>Arabidopsis thaliana</i>; Os, <i>Oryza sativa</i>; Zm, <i>Zea mays</i>), and in gray are fungi (yeast; Sc, <i>Saccharomyces cerevisiae</i>; Sp, <i>Schizosaccharomyces pombe</i>) and mammals (Mm, <i>Mus musculus</i>; Hs, <i>Homo sapiens</i>).</div>
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<img class="figure-thumb" itemprop="thumbnail"
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<strong class="figure-label">
<p> Figure 2. </p>
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<div class="figure-caption-contents"><p> Developmental Abnormalities of <i> ago1 </i> ,… </p></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 2. </p>
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<div class="figure-caption-contents"><p> Developmental Abnormalities of <i> ago1 </i> , <i> fry1 </i> , and <i> xrn </i> Mutants. <b> (A) </b> and <b> (B)… </b> </p></div>
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Figure 2.
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<div class="figure-caption-contents">Developmental Abnormalities of <i>ago1</i>, <i>fry1</i>, and <i>xrn</i> Mutants. <b>(A)</b> and <b>(B)</b> <i>ago1-27</i> and <i>ago1-33</i> mutant lines used for mutagenesis. Plants are dwarf and have serrated leaves. <b>(C)</b> to <b>(F)</b> <i>fry1-4</i> and <i>fry1-5</i> mutants recovered from the mutagenesis screen are shown in the <i>ago1</i> mutant backgrounds (<b>[C]</b> and <b>[D]</b>) or alone (<b>[E]</b> and <b>[F]</b>). Plants are dwarf and have crinkly leaves, rounded leaf margins, and delayed flowering compared with wild-type plants. <b>(G)</b> A Columbia (Col) accession wild-type plant. <b>(H)</b> <i>fry1-6</i> T-DNA insertion mutants display developmental anomalies identical to <i>fry1-4</i> and <i>fry1-5</i> mutants. <b>(I)</b> <i>xrn2-1</i> mutants do not display obvious developmental defects. <b>(K)</b> Hypomorphic <i>xrn3-3</i> mutants display crinkly leaves and delayed flowering. <b>(M)</b> <i>xrn4-6</i> mutants display slightly serrated leaves. <b>(J)</b>, <b>(L)</b>, and <b>(N)</b> <i>xrn2-1 xrn3-3</i> <b>(J)</b>, <i>xrn2-1 xrn4-6</i> <b>(L)</b>, and <i>xrn3-3 xrn4-6</i> <b>(N)</b> double mutants show additive developmental defects, and <i>xrn2-1 xrn3-3</i> mutants <b>(J)</b> resemble <i>fry1-4</i>, <i>fry1-5</i>, and <i>fry1-6</i> single mutants (<b>[E]</b>, <b>[F]</b>, and <b>[H]</b>, respectively).</div>
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<img class="figure-thumb" itemprop="thumbnail"
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<strong class="figure-label">
<p> Figure 3. </p>
</strong>
<div class="figure-caption-contents"><i> ago1 fry1 </i> Double Mutants Posttranscriptionally…</div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 3. </p>
</strong>
<div class="figure-caption-contents"><i> ago1 fry1 </i> Double Mutants Posttranscriptionally Silence L1. RNA gel blot analysis of GUS…</div>
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Figure 3.
</strong>
<div class="figure-caption-contents"><i>ago1 fry1</i> Double Mutants Posttranscriptionally Silence L1. RNA gel blot analysis of GUS mRNA <b>(A)</b> and GUS siRNA <b>(B)</b> accumulation in L1 control and <i>ago1</i>, <i>fry1</i>, and <i>ago1 fry1</i> mutant 18-d-old seedlings. <b>(A)</b> L1 control and L1/<i>fry1-4</i> and L1/<i>fry1-5</i> mutant plants are silenced and accumulate low <i>GUS</i> mRNA levels. <i>ago1-27</i> and <i>ago1-33</i> mutants release L1 silencing and accumulate high levels of <i>GUS</i> mRNA. Both <i>fry1-4</i> and <i>fry1-5</i> restore L1 silencing in <i>ago1</i> mutant backgrounds and accumulate low levels of <i>GUS</i> mRNA, although L1 silencing is restored to a lesser extent in L1/<i>ago1-27/fry1-4</i> mutants (4.2 versus 1.3). Normalized values of GUS mRNA to ethidium bromidestained <i>25S</i> RNA (bottom panel), with L1 levels set at 1.0, are indicated. <b>(B)</b> RNA gel blot analysis of GUS siRNA accumulation. L1 control, L1/<i>fry1-4</i>, L1/<i>fry1-5</i>, L1/<i>ago1-27/fry1-4</i>, and L1/<i>ago1-33/fry1-5</i> mutant plants are silenced and accumulate <i>GUS</i> siRNAs. <i>ago1-27</i> and <i>ago1-33</i> mutants release L1 silencing and do not accumulate detectable <i>GUS</i> siRNAs. Blots were stripped and rehybridized with a probe complementary to <i>U6</i> snRNA as a loading control. Normalized values of GUS siRNAs to U6 small nuclear RNA (snRNA), with L1 levels set at 1.0, are indicated. ND, not detected.</div>
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aria-describedby="figure-caption-3"
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data-image-width="423"
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data-image-alt="Figure 4."
data-pmc-id="PMC2174888"
data-figure-id="fig4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3c9e/2174888/29056e66f198/tpc1903451f04.gif"
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<strong class="figure-label">
<p> Figure 4. </p>
</strong>
<div class="figure-caption-contents"><i> fry1 </i> Mutants Overaccumulate miRNA Target…</div>
</div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Figure 4. </p>
</strong>
<div class="figure-caption-contents"><i> fry1 </i> Mutants Overaccumulate miRNA Target 3 Cleavage Products. RNA gel blot analysis of…</div>
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<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Figure 4.
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<div class="figure-caption-contents"><i>fry1</i> Mutants Overaccumulate miRNA Target 3 Cleavage Products. RNA gel blot analysis of <i>AGO1</i> full-length mRNA and miR168 generated 3 <i>AGO1</i> cleavage products, and <i>ARF10</i>, <i>ARF16</i>, and <i>ARF17</i> full-length RNA and miR160 generated 3 <i>ARF</i> cleavage products in L1 control and <i>ago1</i>, <i>fry1</i>, <i>ago1 fry1</i>, <i>xrn2</i>, <i>xrn3</i>, and <i>xrn4</i> mutant 18-d-old seedlings. <b>(A)</b> <i>AGO1</i> full-length mRNA levels are increased in <i>ago1</i> and <i>ago1/fry1</i> mutants, whereas <i>AGO1</i> 3 cleavage products, but not <i>AGO1</i> full-length mRNA, overaccumulate in <i>fry1</i> single mutants. <b>(B)</b> and <b>(C)</b> <i>fry1</i> mutants and <i>ago1 fry1</i> double mutants overaccumulate <i>ARF10</i>, <i>ARF16</i>, and <i>ARF17</i> 3 cleavage products <b>(B)</b>, similar to <i>xrn4</i> mutants <b>(C)</b>. Ethidium bromidestained <i>25S</i> RNA is shown as a loading control.</div>
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data-pmc-id="PMC2174888"
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<p> Figure 5. </p>
</strong>
<div class="figure-caption-contents"><p> MIRNA Loops Overaccumulate in <i> fry1… </i> </p></div>
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<strong class="figure-label">
<p> Figure 5. </p>
</strong>
<div class="figure-caption-contents"><p> MIRNA Loops Overaccumulate in <i> fry1 </i> and <i> xrn2 xrn3 </i> Double Mutants. <b> (A) </b> Diagram of… </p></div>
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<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Figure 5.
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<div class="figure-caption-contents">MIRNA Loops Overaccumulate in <i>fry1</i> and <i>xrn2 xrn3</i> Double Mutants. <b>(A)</b> Diagram of the various intermediates and end products generated during <i>MIRNA</i> maturation. The stem-loop is the RNA sequence flanked at each end by the miRNA and miRNA* sequences; partial stem-loops are generated after either the 3 end of the miRNA or the 5 end of miRNA* is defined. The positions of the 21-nucleotide miRNA (miR) and miRNA* (miR*) species are indicated by gray boxes (the miR* is the 21-nucleotide RNA species that is opposite of and paired to the miRNA in the <i>MIRNA</i> hairpin). The loop sequence is between the miRNA and the miRNA* and is liberated during miRNA maturation. <b>(B)</b> RNA gel blot analysis of miRNA accumulation in Col control and <i>fry1-4</i> and <i>fry1-5</i> mutant inflorescences. miRNA accumulation is either unchanged or slightly reduced in <i>fry1</i> mutants except for miR168, which slightly overaccumulates. Blots were stripped and rehybridized with a probe complementary to <i>U6</i> RNA as a loading control. <b>(C)</b> RNA gel blot analysis showing that a homozygous <i>fry1-6</i> T-DNA insertion mutant and <i>fry1-4</i> and <i>fry1-5</i> ethyl methanesulfonate mutants overaccumulate <i>MIR168a</i> loops in inflorescence tissues. <i>U6</i> hybridization is shown as a loading control. Normalized values of the MIR168a loop RNA to U6 snRNA, with Col levels set at 1.0, are indicated. <b>(D)</b> RNA gel blot analyses of <i>MIR168a</i> and <i>MIR164b</i> maturation intermediates and end products in inflorescence tissues of controls (Col and <i>MIR168a</i>-overexpressing plants; 35S:168a), hemizygous and homozygous <i>fry1</i> mutants, two homozygous <i>xrn2</i> mutants, a hemizygous <i>xrn3</i> mutant, and a homozygous <i>xrn4</i> mutant. Homozygous <i>fry1</i> mutants overaccumulate <i>MIR168a</i> and <i>MIR164b</i> loops but not stem-loops or partial stem-loops. miR164 and miR168 accumulation in <i>fry1</i> homozygous mutants is shown in <b>(B)</b>. RNA extracted from <i>MIR168a</i>-overexpressing inflorescences was used to indicate the positions of <i>MIR168a</i> stem-loops and partial stem-loops, which are undetectable in wild-type inflorescences. The positions of <sup>32</sup>P-labeled RNA oligonucleotides (nt) are noted. <i>U6</i> hybridization is shown as a loading control. Normalized values of loop RNAs and miRNAs to U6 snRNA, with Col levels set at 1.0, are indicated. <b>(E)</b> RNA gel blot analysis of <i>MIR168a</i> and <i>MIR164b</i> loop accumulation in control Col, <i>xrn2</i>, hypomorphic <i>xrn3</i>, <i>xrn4</i>, <i>xrn2 xrn3</i>, <i>xrn2 xrn4</i>, <i>xrn3 xrn4</i>, and <i>fry1</i> mutant inflorescences. All mutations are homozygous. <i>xrn3</i>-containing mutants overaccumulate loops, although to a lesser extent than <i>fry1</i> mutants. <i>U6</i> hybridization is shown as a loading control. Normalized values of loop RNAs to U6 snRNA, with Col levels set at 1.0, are indicated.</div>
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<p> Figure 6. </p>
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<div class="figure-caption-contents"><p> XRN2 and XRN3 Are Endogenous… </p></div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
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<p> Figure 6. </p>
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<div class="figure-caption-contents"><p> XRN2 and XRN3 Are Endogenous Suppressors of PTGS. GUS protein activities contributed by… </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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Figure 6.
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<div class="figure-caption-contents">XRN2 and XRN3 Are Endogenous Suppressors of PTGS. GUS protein activities contributed by the homozygous transgene loci L1 <b>(A)</b>, Hc1 <b>(B)</b>, and 6b4 <b>(C)</b> in control, <i>ago1</i>, <i>xrn</i>, and <i>fry1</i> mutant leaves. Data are expressed as percentages of silenced plants in each genetic background. The number of individual plants analyzed is indicated above each bar.</div>
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<p> Figure 7. </p>
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<div class="figure-caption-contents"><i> xrn4 </i> and <i> fry1 </i> Mutants Have…</div>
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<p> Figure 7. </p>
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<div class="figure-caption-contents"><i> xrn4 </i> and <i> fry1 </i> Mutants Have Reduced CMV Accumulation. RNA gel blot analyses of…</div>
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Figure 7.
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<div class="figure-caption-contents"><i>xrn4</i> and <i>fry1</i> Mutants Have Reduced CMV Accumulation. RNA gel blot analyses of CMV RNA3 and RNA4 and CMV-derived siRNA accumulation in control L1 and <i>xrn2</i>, <i>xrn3</i>, <i>xrn4</i>, and <i>fry1</i> mutant 29-d-old rosettes. Both <i>fry1</i> and <i>xrn4</i> mutants accumulate less CMV RNA and thus less CMV-derived siRNAs than control L1 plants. 25S rRNA hybridization is shown as a control for mRNA gel blots. Normalized values of CMV RNA3 and RNA4 to <i>25S</i> rRNA, with L1 levels set at 1.0, are indicated. <i>U6</i> hybridization is shown as a loading control for siRNA gel blots. Normalized values of CMV siRNAs to U6 snRNA, with L1 levels set at 1.0, are indicated. The number of symptomatic plants out of 24 CMV-infected plants is indicated at the bottom of each lane.</div>
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<p> Figure 8. </p>
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<div class="figure-caption-contents"><p> Model for FRY1, XRN2, XRN3,… </p></div>
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<p> Figure 8. </p>
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<div class="figure-caption-contents"><p> Model for FRY1, XRN2, XRN3, and XRN4 Activity during PTGS. Shown at left… </p></div>
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Figure 8.
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<div class="figure-caption-contents">Model for FRY1, XRN2, XRN3, and XRN4 Activity during PTGS. Shown at left is a simplified model for miRNA biogenesis and action. Normal miRNA maturation requires DCL1, SERRATE, HYL1, and HEN1 and liberates MIRNA loops, miRNA*, and mature miRNAs. miRNAs are exported from the nucleus by HASTY and incorporate into an AGO1-containing complex to direct the cleavage of partially complementary RNAs. MIRNA loops are substrates of both the nuclear XRN2 and XRN3, while 3 target cleavage products are substrates of the cytoplasmic XRN4. Shown at right is a simplified model for transgene- and virus-induced silencing. Aberrant RNA deriving from transgene transcription and virus replication is converted to double-stranded RNA and processed into siRNAs by RDR6 and SGS3, and DCL2 and DCL4, respectively. siRNAs incorporate into an AGO1-containing complex to direct the cleavage of complementary RNAs. XRN2, XRN3, and XRN4 are endogenous suppressors of PTGS, and we propose that the transgene- and virus-derived aberrant RNAs that trigger PTGS are substrates of these XRNs. At center, FRY1 converts 3-phosphoadenosine 5-phosphate (PAP), a toxic byproduct of sulfate assimilation, into 5 AMP + Pi. Our data, together with those from yeast, predict that XRN levels are regulated through a FRY1-dependent system that keeps PAP, an inhibitor of yeast XRN activity, levels in check. We propose that XRN activities are repressed in <i>fry1</i> mutant plants, leading to the overaccumulation of aberrant RNAs that trigger PTGS.</div>
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