EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair.
Replication fork stalling and collapse is a major source of genome instability leading to neoplastic transformation or cell death. Such stressed replication forks can be conservatively repaired and restarted using homologous recombination (HR) or non-conservatively repaired using micro-homology medi...
Main Authors: | , , , , , , , , , , , , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2015-12-01
|
Series: | PLoS Genetics |
Online Access: | http://europepmc.org/articles/PMC4684289?pdf=render |
id |
doaj-95fc9886b9814bed88fac687edfe81e9 |
---|---|
record_format |
Article |
spelling |
doaj-95fc9886b9814bed88fac687edfe81e92020-11-25T01:52:30ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042015-12-011112e100567510.1371/journal.pgen.1005675EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair.Yuehan WuSuk-Hee LeeElizabeth A WilliamsonBrian L ReinertJu Hwan ChoFen XiaAruna Shanker JaiswalGayathri SrinivasanBhavita PatelAlexis BrantleyDaohong ZhouLijian ShaoRupak PathakMartin Hauer-JensenSudha SinghKimi KongXaiohua WuHyun-Suk KimTimothy BeissbarthJochen GaedckeSandeep BurmaJac A NickoloffRobert A HromasReplication fork stalling and collapse is a major source of genome instability leading to neoplastic transformation or cell death. Such stressed replication forks can be conservatively repaired and restarted using homologous recombination (HR) or non-conservatively repaired using micro-homology mediated end joining (MMEJ). HR repair of stressed forks is initiated by 5' end resection near the fork junction, which permits 3' single strand invasion of a homologous template for fork restart. This 5' end resection also prevents classical non-homologous end-joining (cNHEJ), a competing pathway for DNA double-strand break (DSB) repair. Unopposed NHEJ can cause genome instability during replication stress by abnormally fusing free double strand ends that occur as unstable replication fork repair intermediates. We show here that the previously uncharacterized Exonuclease/Endonuclease/Phosphatase Domain-1 (EEPD1) protein is required for initiating repair and restart of stalled forks. EEPD1 is recruited to stalled forks, enhances 5' DNA end resection, and promotes restart of stalled forks. Interestingly, EEPD1 directs DSB repair away from cNHEJ, and also away from MMEJ, which requires limited end resection for initiation. EEPD1 is also required for proper ATR and CHK1 phosphorylation, and formation of gamma-H2AX, RAD51 and phospho-RPA32 foci. Consistent with a direct role in stalled replication fork cleavage, EEPD1 is a 5' overhang nuclease in an obligate complex with the end resection nuclease Exo1 and BLM. EEPD1 depletion causes nuclear and cytogenetic defects, which are made worse by replication stress. Depleting 53BP1, which slows cNHEJ, fully rescues the nuclear and cytogenetic abnormalities seen with EEPD1 depletion. These data demonstrate that genome stability during replication stress is maintained by EEPD1, which initiates HR and inhibits cNHEJ and MMEJ.http://europepmc.org/articles/PMC4684289?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuehan Wu Suk-Hee Lee Elizabeth A Williamson Brian L Reinert Ju Hwan Cho Fen Xia Aruna Shanker Jaiswal Gayathri Srinivasan Bhavita Patel Alexis Brantley Daohong Zhou Lijian Shao Rupak Pathak Martin Hauer-Jensen Sudha Singh Kimi Kong Xaiohua Wu Hyun-Suk Kim Timothy Beissbarth Jochen Gaedcke Sandeep Burma Jac A Nickoloff Robert A Hromas |
spellingShingle |
Yuehan Wu Suk-Hee Lee Elizabeth A Williamson Brian L Reinert Ju Hwan Cho Fen Xia Aruna Shanker Jaiswal Gayathri Srinivasan Bhavita Patel Alexis Brantley Daohong Zhou Lijian Shao Rupak Pathak Martin Hauer-Jensen Sudha Singh Kimi Kong Xaiohua Wu Hyun-Suk Kim Timothy Beissbarth Jochen Gaedcke Sandeep Burma Jac A Nickoloff Robert A Hromas EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. PLoS Genetics |
author_facet |
Yuehan Wu Suk-Hee Lee Elizabeth A Williamson Brian L Reinert Ju Hwan Cho Fen Xia Aruna Shanker Jaiswal Gayathri Srinivasan Bhavita Patel Alexis Brantley Daohong Zhou Lijian Shao Rupak Pathak Martin Hauer-Jensen Sudha Singh Kimi Kong Xaiohua Wu Hyun-Suk Kim Timothy Beissbarth Jochen Gaedcke Sandeep Burma Jac A Nickoloff Robert A Hromas |
author_sort |
Yuehan Wu |
title |
EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. |
title_short |
EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. |
title_full |
EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. |
title_fullStr |
EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. |
title_full_unstemmed |
EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair. |
title_sort |
eepd1 rescues stressed replication forks and maintains genome stability by promoting end resection and homologous recombination repair. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Genetics |
issn |
1553-7390 1553-7404 |
publishDate |
2015-12-01 |
description |
Replication fork stalling and collapse is a major source of genome instability leading to neoplastic transformation or cell death. Such stressed replication forks can be conservatively repaired and restarted using homologous recombination (HR) or non-conservatively repaired using micro-homology mediated end joining (MMEJ). HR repair of stressed forks is initiated by 5' end resection near the fork junction, which permits 3' single strand invasion of a homologous template for fork restart. This 5' end resection also prevents classical non-homologous end-joining (cNHEJ), a competing pathway for DNA double-strand break (DSB) repair. Unopposed NHEJ can cause genome instability during replication stress by abnormally fusing free double strand ends that occur as unstable replication fork repair intermediates. We show here that the previously uncharacterized Exonuclease/Endonuclease/Phosphatase Domain-1 (EEPD1) protein is required for initiating repair and restart of stalled forks. EEPD1 is recruited to stalled forks, enhances 5' DNA end resection, and promotes restart of stalled forks. Interestingly, EEPD1 directs DSB repair away from cNHEJ, and also away from MMEJ, which requires limited end resection for initiation. EEPD1 is also required for proper ATR and CHK1 phosphorylation, and formation of gamma-H2AX, RAD51 and phospho-RPA32 foci. Consistent with a direct role in stalled replication fork cleavage, EEPD1 is a 5' overhang nuclease in an obligate complex with the end resection nuclease Exo1 and BLM. EEPD1 depletion causes nuclear and cytogenetic defects, which are made worse by replication stress. Depleting 53BP1, which slows cNHEJ, fully rescues the nuclear and cytogenetic abnormalities seen with EEPD1 depletion. These data demonstrate that genome stability during replication stress is maintained by EEPD1, which initiates HR and inhibits cNHEJ and MMEJ. |
url |
http://europepmc.org/articles/PMC4684289?pdf=render |
work_keys_str_mv |
AT yuehanwu eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT sukheelee eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT elizabethawilliamson eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT brianlreinert eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT juhwancho eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT fenxia eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT arunashankerjaiswal eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT gayathrisrinivasan eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT bhavitapatel eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT alexisbrantley eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT daohongzhou eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT lijianshao eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT rupakpathak eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT martinhauerjensen eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT sudhasingh eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT kimikong eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT xaiohuawu eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT hyunsukkim eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT timothybeissbarth eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT jochengaedcke eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT sandeepburma eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT jacanickoloff eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair AT robertahromas eepd1rescuesstressedreplicationforksandmaintainsgenomestabilitybypromotingendresectionandhomologousrecombinationrepair |
_version_ |
1724994788205264896 |