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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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl">
<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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl">
<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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl">
<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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl">
<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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl">
<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="9371772">
<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://www.pnas.org/doi/10.1073/pnas.94.24.12898?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub 0pubmed"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=4487&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=9371772&amp;db=pubmed&amp;nlmid=7505876"
title="See full text options at Atypon"
data-ga-category="full_text"
data-ga-action="Atypon"
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><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--pnas.org-pb-assets-images-pubmed-pnas-pubmed-fulltext.png" alt="Atypon full text link"><span class="text">
Atypon
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/9371772/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=9371772&amp;db=pubmed&amp;nlmid=7505876"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="9371772"
><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="/9371772/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/9371772/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="9371772" 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%2F9371772%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="stFwTEkABTowVbMqWJhg4RAAPt827ZHdh1FJx4QSgV3igfqwN6HdgP5MmXdoeddl"><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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Mammalian capping enzyme complements mutant Saccharomyces cerevisiae lacking mRNA guanylyltransferase and selectively binds the elongating form of RNA polymerase II
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<p>
5'-Capping is an early mRNA modification that has important consequences for downstream events in gene expression. We have isolated mammalian cDNAs encoding capping enzyme. They contain the sequence motifs characteristic of the nucleotidyl transferase superfamily. The predicted mouse and human enzymes consist of 597 amino acids and are 95% identical. Mouse cDNA directed synthesis of a guanylylated 68-kDa polypeptide that also contained RNA 5'-triphosphatase activity and catalyzed formation of RNA 5'-terminal GpppG. A haploid strain of Saccharomyces cerevisiae lacking mRNA guanylyltransferase was complemented for growth by the mouse cDNA. Conversion of Lys-294 in the KXDG-conserved motif eliminated both guanylylation and complementation, identifying it as the active site. The K294A mutant retained RNA 5'-triphosphatase activity, which was eliminated by N-terminal truncation. Full-length capping enzyme and an active C-terminal fragment bound to the elongating form and not to the initiating form of polymerase. The results document functional conservation of eukaryotic mRNA guanylyltransferases from yeast to mammals and indicate that the phosphorylated C-terminal domain of RNA polymerase II couples capping to transcription elongation. These results also explain the selective capping of RNA polymerase II transcripts.
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<p> Figure 1 </p>
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<div class="figure-caption-contents"><p> MCE and human capping enzyme… </p></div>
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<p> Figure 1 </p>
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<div class="figure-caption-contents"><p> MCE and human capping enzyme cDNA clones. ( <i> A </i> ) Schematic diagram of… </p></div>
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Figure 1
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<div class="figure-caption-contents">MCE and human capping enzyme cDNA clones. (<i>A</i>) Schematic diagram of the ORFs (1,794 bp, solid bars), relative sizes of the untranslated regions, and the sequences and positions of initiation sites and polyadenylation signals. (<i>B</i>) Predicted amino acid sequences of the mouse and human enzymes. Residues that are identical (95% of total) are omitted from the humansequence.</div>
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<p> Figure 2 </p>
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<div class="figure-caption-contents"><p> Sequence alignments of cellular mRNA… </p></div>
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<p> Figure 2 </p>
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<div class="figure-caption-contents"><p> Sequence alignments of cellular mRNA guanylyltransferases. The deduced amino sequences of the C-terminal… </p></div>
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Figure 2
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<div class="figure-caption-contents">Sequence alignments of cellular mRNA guanylyltransferases. The deduced amino sequences of the C-terminal regions of the mouse and human enzymes (residues 232597) are compared with <i>C. elegans CEL-1</i> (residues 238573; ref. 19) and to the nearly full-length <i>S. pombe</i> (13) and <i>S. cerevisiae</i> (9) guanylyltransferase subunits of capping enzyme. Dark background indicates regions of sequence identity; the active site motif, including Lys-294, in the mammalian enzymes isbracketed.</div>
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<p> Figure 3 </p>
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<div class="figure-caption-contents"><p> Expression of capping enzyme transcripts… </p></div>
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<p> Figure 3 </p>
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<div class="figure-caption-contents"><p> Expression of capping enzyme transcripts <i> in vivo </i> . ( <i> A </i> ) Poly(A) <sup> +… </sup> </p></div>
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Figure 3
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<div class="figure-caption-contents">Expression of capping enzyme transcripts <i>in vivo</i>. (<i>A</i>) Poly(A)<sup>+</sup> RNAs prepared from mouse adult tissues (<i>Left</i>) and embryos at the indicated days of gestation (<i>Right</i>) were probed with a mouse EST as described in <i>Materials and Methods</i>. (<i>B</i>) Human adult tissue poly(A)<sup>+</sup> RNAs were probed with the same EST and a human β-actin cDNA. Positions of marker RNAs and hybridized transcripts (arrows) areindicated.</div>
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<p> Figure 4 </p>
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<div class="figure-caption-contents"><i> In vitro </i> synthesis and specific…</div>
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<p> Figure 4 </p>
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<div class="figure-caption-contents"><i> In vitro </i> synthesis and specific immunoprecipitation of MCEs. ( <i> A </i> ) <sup> 35 </sup> S-methionine-labeled…</div>
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Figure 4
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<div class="figure-caption-contents"><i>In vitro</i> synthesis and specific immunoprecipitation of MCEs. (<i>A</i>) <sup>35</sup>S-methionine-labeled proteins were synthesized <i>in vitro</i> by using luciferase (Luc) cDNA (lane 1) or cDNAs for wild type (lane 2), mutant (lane 3), and C-terminal fragment (Δ210, lane 4) of MCE. Products were analyzed by SDS/PAGE and autoradiography. (<i>B</i>) Products shown in <i>A</i> were immunoprecipitated with αMCE and analyzed by SDS/PAGE andautoradiography.</div>
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<img class="figure-thumb" itemprop="thumbnail"
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<p> Figure 5 </p>
</strong>
<div class="figure-caption-contents"><p> Guanylyltransferase activity of MCEs. (… </p></div>
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<p> Figure 5 </p>
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<div class="figure-caption-contents"><p> Guanylyltransferase activity of MCEs. ( <i> A </i> ) <i> In vitro </i> translated and immunoprecipitated proteins… </p></div>
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Figure 5
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<div class="figure-caption-contents">Guanylyltransferase activity of MCEs. (<i>A</i>) <i>In vitro</i> translated and immunoprecipitated proteins were incubated with α[<sup>32</sup>P]GTP as described in <i>Materials and Methods</i> and analyzed by SDS/PAGE and autoradiography. Lane 1, no added DNA; lanes 25, products synthesized with cDNA templates for full-length MCE, C-terminal fragment (Δ210), and mutants K294A and K290A, respectively. Mouse L cell nuclear extract (lane 6) and partially purified HeLa cell capping enzyme (lane 7) were immunoprecipitated with αMCE and then incubated with α[<sup>32</sup>P]GTP. Proteins were analyzed by SDS/PAGE and autoradiography. (<i>B</i>) 5-Terminal guanylylation of T7 transcripts was assayed by incubating the RNA with α[<sup>32</sup>P]GTP and immunoprecipitates of <i>in vitro</i> synthesized wild-type (lane 2) or K294A mutant (lane 3) MCE or vaccinia virus capping enzyme (VCE, lane 4). (<i>C</i>) Cap formation on the radiolabeled RNAs shown in <i>B</i> was verified by digestion with P1 nuclease followed by alkaline phosphatase and TLC analysis of the digests with authentic GpppG and GppppG asmarkers.</div>
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<p> Figure 6 </p>
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<div class="figure-caption-contents"><p> MCE contains RNA 5-triphosphatase. RNA… </p></div>
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<p> Figure 6 </p>
</strong>
<div class="figure-caption-contents"><p> MCE contains RNA 5-triphosphatase. RNA containing 5-terminal γ[ <sup> 32 </sup> P]GTP, prepared as described… </p></div>
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Figure 6
</strong>
<div class="figure-caption-contents">MCE contains RNA 5-triphosphatase. RNA containing 5-terminal γ[<sup>32</sup>P]GTP, prepared as described in <i>Materials and Methods</i>, was incubated with <i>in vitro</i>-synthesized, immunoprecipitated MCE wild type (lane 2), mutant K294A (lane 3), N-terminal truncation mutants Δ210 (lane 4), and Δ144 (lane 5) or vaccinia virus capping enzyme (lane 1). Samples were analyzed for release of labeled Pi by TLC andautoradiography.</div>
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<p> Figure 7 </p>
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<div class="figure-caption-contents"><p> Complementation of <i> S. cerevisiae </i> lacking… </p></div>
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<p> Figure 7 </p>
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<div class="figure-caption-contents"><p> Complementation of <i> S. cerevisiae </i> lacking mRNA guanylyltransferase. Growth was measured on selective media… </p></div>
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Figure 7
</strong>
<div class="figure-caption-contents">Complementation of <i>S. cerevisiae</i> lacking mRNA guanylyltransferase. Growth was measured on selective media using <i>CEG1</i><sup></sup> cells transformed with empty parental pG-1 (vector), pG-1 containing wild-type (MCE) or mutant (K294A) MCE or with a <i>CEG1</i>-containing <i>URA3</i> plasmid (<i>CEG1</i>) as detailed in <i>Materials andMethods</i>.</div>
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<p> Figure 8 </p>
</strong>
<div class="figure-caption-contents"><p> Capping enzyme directly interacts with… </p></div>
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<p> Figure 8 </p>
</strong>
<div class="figure-caption-contents"><p> Capping enzyme directly interacts with the phosphorylated form of RNA pol II. (… </p></div>
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Figure 8
</strong>
<div class="figure-caption-contents">Capping enzyme directly interacts with the phosphorylated form of RNA pol II. (<i>A</i>) A HeLa cell protein fraction containing approximately equal amounts of the phosphorylated (pol IIo) and unphosphorylated (pol IIa) forms of pol II (lane 1) was mixed with αMCE-protein A beads in the presence or absence of affinity-purified HeLa capping enzyme (HCE, lane 2). Immunoprecipitates (lane 3 and 4) were extensively washed and loaded onto a 520% gradient SDS-polyacrylamide gel with the input (10%, lanes 1 and 2) for Western blot analysis. Immunoblots were developed for the CTD of the largest subunit of pol II (<i>Upper</i>) or capping enzyme (<i>Lower</i>). (<i>B</i>) The carboxy-terminal domain of capping enzyme is sufficient to mediate direct and selective interaction with the phosphorylated form of pol II. The HeLa protein fraction containing the two forms of pol II (lane 1) was mixed with antihexahistidine tag antibody (α6xHis)-protein G beads in the presence or absence of purified, recombinant hexa-histidine tagged carboxy-terminal domain of MCE Δ210. Immunoprecipitates (lanes 2 and 3) were extensively washed and loaded onto a 520% gradient SDS-polyacrylamide gel with the input (10%, lane 1) for Western blot analysis. Immunoblots were developed for the CTD of the largest subunit of pol II (<i>Upper</i>) or MCE(<i>Lower</i>).</div>
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Origins of mRNA identity: capping enzymes bind to the phosphorylated C-terminal domain of RNA polymerase II.
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