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| Status |
Public on Oct 17, 2016 |
| Title |
Pif1-family helicases cooperate to suppress widespread replication fork arrest at tRNA genes |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Other
|
| Summary |
Saccharomyces cerevisiae encodes two distinct Pif1-family helicases – Pif1 and Rrm3 – which have been reported to play distinct roles in numerous nuclear processes. Here, we systematically characterize the roles of Pif1 helicases in replisome progression and lagging- strand synthesis in S. cerevisiae. We demonstrate that either Pif1 or Rrm3 redundantly stimulate strand-displacement by DNA polymerase δ during lagging-strand synthesis. By analyzing replisome mobility in pif1 and rrm3 mutants, we show that Rrm3, with a partially redundant contribution from Pif1, suppresses widespread terminal arrest of the replisome at tRNA genes. Although both head-on and codirectional collisions induce replication fork arrest at tRNA genes, head-on collisions arrest a higher proportion of replisomes; consistent with this observation, we find that head-on collisions between tRNA transcription and replisome progression are under-represented in the S. cerevisiae genome. Further, we demonstrate that tRNA-mediated arrest is R-loop independent, and propose that replisome arrest and DNA damage are mechanistically separable.
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| Overall design |
We enriched Okazaki fragments by depletion of DNA ligase I, and sequenced the resulting enriched single-stranded/nicked DNA
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| Contributor(s) |
Osmundson JS, Kumar J, Yeung R, Smith DJ |
| Citation(s) |
27991904 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 GM114340 |
Direct in vivo characterization of eukaryotic replisome activity |
NEW YORK UNIVERSITY |
Smith |
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| |
| Submission date |
Aug 12, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Duncan Smith |
| E-mail(s) |
duncan.smith@nyu.edu
|
| Phone |
212-992-6595
|
| Organization name |
New York University
|
| Department |
Dept. of Biology/Center for Genomics and Systems Biology
|
| Street address |
1009 Silver Center, 100 Washington Sq E
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10003 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (41)
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| Relations |
| BioProject |
PRJNA292617 |
| SRA |
SRP062301 |
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