Whole genome RNAi screens reveal a critical role of REV3 in coping with replication stress.

Details

Serval ID
serval:BIB_97F709D9E124
Type
Article: article from journal or magazin.
Collection
Publications
Title
Whole genome RNAi screens reveal a critical role of REV3 in coping with replication stress.
Journal
Molecular Oncology
Author(s)
Kotov I.N., Siebring-van Olst E., Knobel P.A., van der Meulen-Muileman I.H., Felley-Bosco E., van Beusechem V.W., Smit E.F., Stahel R.A., Marti T.M.
ISSN
1878-0261 (Electronic)
ISSN-L
1574-7891
Publication state
Published
Issued date
2014
Volume
8
Number
8
Pages
1747-1759
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Abstract
REV3, the catalytic subunit of translesion polymerase zeta (polζ), is commonly associated with DNA damage bypass and repair. Despite sharing accessory subunits with replicative polymerase δ, very little is known about the role of polζ in DNA replication. We previously demonstrated that inhibition of REV3 expression induces persistent DNA damage and growth arrest in cancer cells. To reveal determinants of this sensitivity and obtain insights into the cellular function of REV3, we performed whole human genome RNAi library screens aimed at identification of synthetic lethal interactions with REV3 in A549 lung cancer cells. The top confirmed hit was RRM1, the large subunit of ribonucleotide reductase (RNR), a critical enzyme of de novo nucleotide synthesis. Treatment with the RNR-inhibitor hydroxyurea (HU) synergistically increased the fraction of REV3-deficient cells containing single stranded DNA (ssDNA) as indicated by an increase in replication protein A (RPA). However, this increase was not accompanied by accumulation of the DNA damage marker γH2AX suggesting a role of REV3 in counteracting HU-induced replication stress (RS). Consistent with a role of REV3 in DNA replication, increased RPA staining was confined to HU-treated S-phase cells. Additionally, we found genes related to RS to be significantly enriched among the top hits of the synthetic sickness/lethality (SSL) screen further corroborating the importance of REV3 for DNA replication under conditions of RS.
Keywords
Cell Line, Tumor, DNA Replication/genetics, DNA Replication/physiology, DNA-Binding Proteins/genetics, DNA-Binding Proteins/metabolism, DNA-Directed DNA Polymerase/genetics, DNA-Directed DNA Polymerase/metabolism, Flow Cytometry, Humans, RNA Interference/physiology, Tumor Suppressor Proteins/genetics, Tumor Suppressor Proteins/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
26/01/2015 13:00
Last modification date
20/08/2019 14:59
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