Loading…

Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p

Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 2009-02, Vol.37 (2), p.354-367
Main Authors: Zappulla, David C, Roberts, Jennifer N, Goodrich, Karen J, Cech, Thomas R, Wuttke, Deborah S
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3
cites cdi_FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3
container_end_page 367
container_issue 2
container_start_page 354
container_title Nucleic acids research
container_volume 37
creator Zappulla, David C
Roberts, Jennifer N
Goodrich, Karen J
Cech, Thomas R
Wuttke, Deborah S
description Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified Cdc13p on telomerase action in vitro. We show that the full-length protein and its DNA-binding domain (DBD) inhibit primer extension by telomerase. This inhibition occurs by competitive blocking of telomerase access to DNA. To further understand the requirements for productive telomerase 3'-end access when Cdc13p or the DBD is bound to a telomerase substrate, we constrained protein binding at various distances from the 3'-end on two sets of increasingly longer oligonucleotides. We find that Cdc13p inhibits the action of telomerase through three distinct biochemical modes, including inhibiting telomerase even when a significant tail is available, representing a novel 'action at a distance' inhibitory activity. Thus, while yeast Cdc13p exhibits the same general activity as human POT1, providing an off switch for telomerase when bound near the 3'-end, there are significant mechanistic differences in the ways telomere end-binding proteins inhibit telomerase action.
doi_str_mv 10.1093/nar/gkn830
format article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2632905</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><oup_id>10.1093/nar/gkn830</oup_id><sourcerecordid>20405213</sourcerecordid><originalsourceid>FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3</originalsourceid><addsrcrecordid>eNqF0U-P1CAYBnBiNO64evEDaGOiB2PdFygULibrrDqbbPTgrjFeCAU6w24HKrTG-fZWO65_Dnri8PzyBHgQuo_hOQZJj4JOR-urICjcQAtMOSkryclNtAAKrMRQiQN0J-dLAFxhVt1GB1hCRaGuFuj8NGx84wcfQxHbYud0HorBdXHrks6u0OZH1OyKYeN-Bt4UOZ-8PS4bH6wP66JPcXA-PCuW1mDa30W3Wt1ld29_HqKL16_Ol6vy7N2b0-XxWWkY4UNJqTONJo2wFXOEGStcba1zRhLONWOE4sbQFoygvLXcNlIYWROtJbYE6pYeohdzbz82W2eNC0PSneqT3-q0U1F79WcS_Eat4xdFOCUS2FTwZF-Q4ufR5UFtfTau63RwccyKc8E5ZdV_IYEKGMF0go_-gpdxTGH6hckAx4wLMaGnMzIp5pxce31lDOr7pGqaVM2TTvjB74_8RfcbTuDxDOLY_7uonJ3Pg_t6LXW6UrymNVOrj58UebkiH5gk6mTyD2ff6qj0OvmsLt4TwBQwEzXUmH4DZJfCvQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>200615688</pqid></control><display><type>article</type><title>Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p</title><source>Open Access: Oxford University Press Open Journals</source><source>PubMed Central</source><creator>Zappulla, David C ; Roberts, Jennifer N ; Goodrich, Karen J ; Cech, Thomas R ; Wuttke, Deborah S</creator><creatorcontrib>Zappulla, David C ; Roberts, Jennifer N ; Goodrich, Karen J ; Cech, Thomas R ; Wuttke, Deborah S</creatorcontrib><description>Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified Cdc13p on telomerase action in vitro. We show that the full-length protein and its DNA-binding domain (DBD) inhibit primer extension by telomerase. This inhibition occurs by competitive blocking of telomerase access to DNA. To further understand the requirements for productive telomerase 3'-end access when Cdc13p or the DBD is bound to a telomerase substrate, we constrained protein binding at various distances from the 3'-end on two sets of increasingly longer oligonucleotides. We find that Cdc13p inhibits the action of telomerase through three distinct biochemical modes, including inhibiting telomerase even when a significant tail is available, representing a novel 'action at a distance' inhibitory activity. Thus, while yeast Cdc13p exhibits the same general activity as human POT1, providing an off switch for telomerase when bound near the 3'-end, there are significant mechanistic differences in the ways telomere end-binding proteins inhibit telomerase action.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gkn830</identifier><identifier>PMID: 19043074</identifier><identifier>CODEN: NARHAD</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Nuclease Protection Assays ; Nucleic Acid Enzymes ; Protein Structure, Tertiary ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - enzymology ; Saccharomyces cerevisiae Proteins - chemistry ; Saccharomyces cerevisiae Proteins - metabolism ; Telomerase - antagonists &amp; inhibitors ; Telomere - metabolism ; Telomere-Binding Proteins - chemistry ; Telomere-Binding Proteins - metabolism</subject><ispartof>Nucleic acids research, 2009-02, Vol.37 (2), p.354-367</ispartof><rights>2008 The Author(s) 2008</rights><rights>2008 The Author(s)</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3</citedby><cites>FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632905/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2632905/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,1604,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19043074$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zappulla, David C</creatorcontrib><creatorcontrib>Roberts, Jennifer N</creatorcontrib><creatorcontrib>Goodrich, Karen J</creatorcontrib><creatorcontrib>Cech, Thomas R</creatorcontrib><creatorcontrib>Wuttke, Deborah S</creatorcontrib><title>Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified Cdc13p on telomerase action in vitro. We show that the full-length protein and its DNA-binding domain (DBD) inhibit primer extension by telomerase. This inhibition occurs by competitive blocking of telomerase access to DNA. To further understand the requirements for productive telomerase 3'-end access when Cdc13p or the DBD is bound to a telomerase substrate, we constrained protein binding at various distances from the 3'-end on two sets of increasingly longer oligonucleotides. We find that Cdc13p inhibits the action of telomerase through three distinct biochemical modes, including inhibiting telomerase even when a significant tail is available, representing a novel 'action at a distance' inhibitory activity. Thus, while yeast Cdc13p exhibits the same general activity as human POT1, providing an off switch for telomerase when bound near the 3'-end, there are significant mechanistic differences in the ways telomere end-binding proteins inhibit telomerase action.</description><subject>Nuclease Protection Assays</subject><subject>Nucleic Acid Enzymes</subject><subject>Protein Structure, Tertiary</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - enzymology</subject><subject>Saccharomyces cerevisiae Proteins - chemistry</subject><subject>Saccharomyces cerevisiae Proteins - metabolism</subject><subject>Telomerase - antagonists &amp; inhibitors</subject><subject>Telomere - metabolism</subject><subject>Telomere-Binding Proteins - chemistry</subject><subject>Telomere-Binding Proteins - metabolism</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>TOX</sourceid><recordid>eNqF0U-P1CAYBnBiNO64evEDaGOiB2PdFygULibrrDqbbPTgrjFeCAU6w24HKrTG-fZWO65_Dnri8PzyBHgQuo_hOQZJj4JOR-urICjcQAtMOSkryclNtAAKrMRQiQN0J-dLAFxhVt1GB1hCRaGuFuj8NGx84wcfQxHbYud0HorBdXHrks6u0OZH1OyKYeN-Bt4UOZ-8PS4bH6wP66JPcXA-PCuW1mDa30W3Wt1ld29_HqKL16_Ol6vy7N2b0-XxWWkY4UNJqTONJo2wFXOEGStcba1zRhLONWOE4sbQFoygvLXcNlIYWROtJbYE6pYeohdzbz82W2eNC0PSneqT3-q0U1F79WcS_Eat4xdFOCUS2FTwZF-Q4ufR5UFtfTau63RwccyKc8E5ZdV_IYEKGMF0go_-gpdxTGH6hckAx4wLMaGnMzIp5pxce31lDOr7pGqaVM2TTvjB74_8RfcbTuDxDOLY_7uonJ3Pg_t6LXW6UrymNVOrj58UebkiH5gk6mTyD2ff6qj0OvmsLt4TwBQwEzXUmH4DZJfCvQ</recordid><startdate>20090201</startdate><enddate>20090201</enddate><creator>Zappulla, David C</creator><creator>Roberts, Jennifer N</creator><creator>Goodrich, Karen J</creator><creator>Cech, Thomas R</creator><creator>Wuttke, Deborah S</creator><general>Oxford University Press</general><general>Oxford Publishing Limited (England)</general><scope>FBQ</scope><scope>BSCLL</scope><scope>TOX</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7QP</scope><scope>7QR</scope><scope>7SS</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20090201</creationdate><title>Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p</title><author>Zappulla, David C ; Roberts, Jennifer N ; Goodrich, Karen J ; Cech, Thomas R ; Wuttke, Deborah S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Nuclease Protection Assays</topic><topic>Nucleic Acid Enzymes</topic><topic>Protein Structure, Tertiary</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - enzymology</topic><topic>Saccharomyces cerevisiae Proteins - chemistry</topic><topic>Saccharomyces cerevisiae Proteins - metabolism</topic><topic>Telomerase - antagonists &amp; inhibitors</topic><topic>Telomere - metabolism</topic><topic>Telomere-Binding Proteins - chemistry</topic><topic>Telomere-Binding Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zappulla, David C</creatorcontrib><creatorcontrib>Roberts, Jennifer N</creatorcontrib><creatorcontrib>Goodrich, Karen J</creatorcontrib><creatorcontrib>Cech, Thomas R</creatorcontrib><creatorcontrib>Wuttke, Deborah S</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Open Access: Oxford University Press Open Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Calcium &amp; Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health &amp; Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zappulla, David C</au><au>Roberts, Jennifer N</au><au>Goodrich, Karen J</au><au>Cech, Thomas R</au><au>Wuttke, Deborah S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2009-02-01</date><risdate>2009</risdate><volume>37</volume><issue>2</issue><spage>354</spage><epage>367</epage><pages>354-367</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><coden>NARHAD</coden><abstract>Appropriate control of the chromosome end-replicating enzyme telomerase is crucial for maintaining telomere length and genomic stability. The essential telomeric DNA-binding protein Cdc13p both positively and negatively regulates telomere length in budding yeast. Here we test the effect of purified Cdc13p on telomerase action in vitro. We show that the full-length protein and its DNA-binding domain (DBD) inhibit primer extension by telomerase. This inhibition occurs by competitive blocking of telomerase access to DNA. To further understand the requirements for productive telomerase 3'-end access when Cdc13p or the DBD is bound to a telomerase substrate, we constrained protein binding at various distances from the 3'-end on two sets of increasingly longer oligonucleotides. We find that Cdc13p inhibits the action of telomerase through three distinct biochemical modes, including inhibiting telomerase even when a significant tail is available, representing a novel 'action at a distance' inhibitory activity. Thus, while yeast Cdc13p exhibits the same general activity as human POT1, providing an off switch for telomerase when bound near the 3'-end, there are significant mechanistic differences in the ways telomere end-binding proteins inhibit telomerase action.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>19043074</pmid><doi>10.1093/nar/gkn830</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0305-1048
ispartof Nucleic acids research, 2009-02, Vol.37 (2), p.354-367
issn 0305-1048
1362-4962
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2632905
source Open Access: Oxford University Press Open Journals; PubMed Central
subjects Nuclease Protection Assays
Nucleic Acid Enzymes
Protein Structure, Tertiary
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae Proteins - chemistry
Saccharomyces cerevisiae Proteins - metabolism
Telomerase - antagonists & inhibitors
Telomere - metabolism
Telomere-Binding Proteins - chemistry
Telomere-Binding Proteins - metabolism
title Inhibition of yeast telomerase action by the telomeric ssDNA-binding protein, Cdc13p
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T09%3A27%3A20IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Inhibition%20of%20yeast%20telomerase%20action%20by%20the%20telomeric%20ssDNA-binding%20protein,%20Cdc13p&rft.jtitle=Nucleic%20acids%20research&rft.au=Zappulla,%20David%20C&rft.date=2009-02-01&rft.volume=37&rft.issue=2&rft.spage=354&rft.epage=367&rft.pages=354-367&rft.issn=0305-1048&rft.eissn=1362-4962&rft.coden=NARHAD&rft_id=info:doi/10.1093/nar/gkn830&rft_dat=%3Cproquest_pubme%3E20405213%3C/proquest_pubme%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c526t-33ecba2b8d45e25cd8e7ddeec9266a55231bc3f0c836fd6db98c972aa91d207f3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=200615688&rft_id=info:pmid/19043074&rft_oup_id=10.1093/nar/gkn830&rfr_iscdi=true