The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres.
Telomerase, the enzyme that maintains telomeres, preferentially lengthens short telomeres. The S. cerevisiae Pif1 DNA helicase inhibits both telomerase-mediated telomere lengthening and de novo telomere addition at double strand breaks (DSB). Here, we report that the association of the telomerase su...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-04-01
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| Series: | PLoS Genetics |
| Online Access: | http://europepmc.org/articles/PMC4408051?pdf=render |
| _version_ | 1819050603536449536 |
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| author | Jane A Phillips Angela Chan Katrin Paeschke Virginia A Zakian |
| author_facet | Jane A Phillips Angela Chan Katrin Paeschke Virginia A Zakian |
| author_sort | Jane A Phillips |
| collection | DOAJ |
| description | Telomerase, the enzyme that maintains telomeres, preferentially lengthens short telomeres. The S. cerevisiae Pif1 DNA helicase inhibits both telomerase-mediated telomere lengthening and de novo telomere addition at double strand breaks (DSB). Here, we report that the association of the telomerase subunits Est2 and Est1 at a DSB was increased in the absence of Pif1, as it is at telomeres, suggesting that Pif1 suppresses de novo telomere addition by removing telomerase from the break. To determine how the absence of Pif1 results in telomere lengthening, we used the single telomere extension assay (STEX), which monitors lengthening of individual telomeres in a single cell cycle. In the absence of Pif1, telomerase added significantly more telomeric DNA, an average of 72 nucleotides per telomere compared to the 45 nucleotides in wild type cells, and the fraction of telomeres lengthened increased almost four-fold. Using an inducible short telomere assay, Est2 and Est1 no longer bound preferentially to a short telomere in pif1 mutant cells while binding of Yku80, a telomere structural protein, was unaffected by the status of the PIF1 locus. Two experiments demonstrate that Pif1 binding is affected by telomere length: Pif1 (but not Yku80) -associated telomeres were 70 bps longer than bulk telomeres, and in the inducible short telomere assay, Pif1 bound better to wild type length telomeres than to short telomeres. Thus, preferential lengthening of short yeast telomeres is achieved in part by targeting the negative regulator Pif1 to long telomeres. |
| first_indexed | 2024-12-21T11:50:40Z |
| format | Article |
| id | doaj.art-2ece023f2dec4b06b492f689739de9c5 |
| institution | Directory Open Access Journal |
| issn | 1553-7390 1553-7404 |
| language | English |
| last_indexed | 2024-12-21T11:50:40Z |
| publishDate | 2015-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj.art-2ece023f2dec4b06b492f689739de9c52022-12-21T19:05:03ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042015-04-01114e100518610.1371/journal.pgen.1005186The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres.Jane A PhillipsAngela ChanKatrin PaeschkeVirginia A ZakianTelomerase, the enzyme that maintains telomeres, preferentially lengthens short telomeres. The S. cerevisiae Pif1 DNA helicase inhibits both telomerase-mediated telomere lengthening and de novo telomere addition at double strand breaks (DSB). Here, we report that the association of the telomerase subunits Est2 and Est1 at a DSB was increased in the absence of Pif1, as it is at telomeres, suggesting that Pif1 suppresses de novo telomere addition by removing telomerase from the break. To determine how the absence of Pif1 results in telomere lengthening, we used the single telomere extension assay (STEX), which monitors lengthening of individual telomeres in a single cell cycle. In the absence of Pif1, telomerase added significantly more telomeric DNA, an average of 72 nucleotides per telomere compared to the 45 nucleotides in wild type cells, and the fraction of telomeres lengthened increased almost four-fold. Using an inducible short telomere assay, Est2 and Est1 no longer bound preferentially to a short telomere in pif1 mutant cells while binding of Yku80, a telomere structural protein, was unaffected by the status of the PIF1 locus. Two experiments demonstrate that Pif1 binding is affected by telomere length: Pif1 (but not Yku80) -associated telomeres were 70 bps longer than bulk telomeres, and in the inducible short telomere assay, Pif1 bound better to wild type length telomeres than to short telomeres. Thus, preferential lengthening of short yeast telomeres is achieved in part by targeting the negative regulator Pif1 to long telomeres.http://europepmc.org/articles/PMC4408051?pdf=render |
| spellingShingle | Jane A Phillips Angela Chan Katrin Paeschke Virginia A Zakian The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. PLoS Genetics |
| title | The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. |
| title_full | The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. |
| title_fullStr | The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. |
| title_full_unstemmed | The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. |
| title_short | The pif1 helicase, a negative regulator of telomerase, acts preferentially at long telomeres. |
| title_sort | pif1 helicase a negative regulator of telomerase acts preferentially at long telomeres |
| url | http://europepmc.org/articles/PMC4408051?pdf=render |
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