Integrator facilitates RNAPII removal to prevent transcription-replication collisions and genome instability

Rahul Bhowmick; Kavi P.M. Mehta; Mads Lerdrup; David Cortez

doi:10.1016/j.molcel.2023.05.015

Integrator facilitates RNAPII removal to prevent transcription-replication collisions and genome instability

Rahul Bhowmick^*, Kavi P.M. Mehta, Mads Lerdrup, David Cortez

^*Corresponding author af dette arbejde

Molecular Aging Program

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

11 Citationer (Scopus)

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Abstract

DNA replication preferentially initiates close to active transcription start sites (TSSs) in the human genome. Transcription proceeds discontinuously with an accumulation of RNA polymerase II (RNAPII) in a paused state near the TSS. Consequently, replication forks inevitably encounter paused RNAPII soon after replication initiates. Hence, dedicated machinery may be needed to remove RNAPII and facilitate unperturbed fork progression. In this study, we discovered that Integrator, a transcription termination machinery involved in the processing of RNAPII transcripts, interacts with the replicative helicase at active forks and promotes the removal of RNAPII from the path of the replication fork. Integrator-deficient cells have impaired replication fork progression and accumulate hallmarks of genome instability including chromosome breaks and micronuclei. The Integrator complex resolves co-directional transcription-replication conflicts to facilitate faithful DNA replication.

Originalsprog	Engelsk
Tidsskrift	Molecular Cell
Vol/bind	83
Udgave nummer	13
Sider (fra-til)	2357-2366.e8
ISSN	1097-2765
DOI	https://doi.org/10.1016/j.molcel.2023.05.015
Status	Udgivet - 2023

Bibliografisk note

Funding Information:
We thank Karen Adelman (Harvard University), Chad Stein (Harvard University), and Karlene Cimprich (Stanford University) for providing INTS11 degron cells and HO/CD cell lines and helpful discussions. We thank Ian Hickson (Copenhagen University) and Eva Petermann (University of Birmingham) for providing RNaseH1 and TBP overexpressing plasmids. This research was supported by the Breast Cancer Research Foundation , NIH grant R01CA239161 , and a pilot project supported by Vanderbilt-Ingram Cancer Center grant P30CA068485 to D.C. K.P.M.M. is supported by K99ES034058 .

Funding Information:
We thank Karen Adelman (Harvard University), Chad Stein (Harvard University), and Karlene Cimprich (Stanford University) for providing INTS11 degron cells and HO/CD cell lines and helpful discussions. We thank Ian Hickson (Copenhagen University) and Eva Petermann (University of Birmingham) for providing RNaseH1 and TBP overexpressing plasmids. This research was supported by the Breast Cancer Research Foundation, NIH grant R01CA239161, and a pilot project supported by Vanderbilt-Ingram Cancer Center grant P30CA068485 to D.C. K.P.M.M. is supported by K99ES034058. D.C and R.B. conceived the project and designed the experiments. R.B. and K.P.M.M. performed the experiments. D.C. R.B. and K.P.M.M. analyzed the data. M.L. performed all the bioinformatic analysis. D.C. and R.B. wrote the manuscript, and all authors edited it. D.C. and R.B. supervised the project. The authors declare no competing interests.

Publisher Copyright:
© 2023 Elsevier Inc.

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10.1016/j.molcel.2023.05.015

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Citationsformater

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title = "Integrator facilitates RNAPII removal to prevent transcription-replication collisions and genome instability",

abstract = "DNA replication preferentially initiates close to active transcription start sites (TSSs) in the human genome. Transcription proceeds discontinuously with an accumulation of RNA polymerase II (RNAPII) in a paused state near the TSS. Consequently, replication forks inevitably encounter paused RNAPII soon after replication initiates. Hence, dedicated machinery may be needed to remove RNAPII and facilitate unperturbed fork progression. In this study, we discovered that Integrator, a transcription termination machinery involved in the processing of RNAPII transcripts, interacts with the replicative helicase at active forks and promotes the removal of RNAPII from the path of the replication fork. Integrator-deficient cells have impaired replication fork progression and accumulate hallmarks of genome instability including chromosome breaks and micronuclei. The Integrator complex resolves co-directional transcription-replication conflicts to facilitate faithful DNA replication.",

keywords = "fork reversal, genome stability, Integrator complex, MCM complex, R-loop, replication stress, replicative helicase, RNA polymerase II, RNAPII, transcription-replication conflicts, TRCs",

author = "Rahul Bhowmick and Mehta, {Kavi P.M.} and Mads Lerdrup and David Cortez",

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year = "2023",

doi = "10.1016/j.molcel.2023.05.015",

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TY - JOUR

T1 - Integrator facilitates RNAPII removal to prevent transcription-replication collisions and genome instability

AU - Bhowmick, Rahul

AU - Mehta, Kavi P.M.

AU - Lerdrup, Mads

AU - Cortez, David

PY - 2023

Y1 - 2023

N2 - DNA replication preferentially initiates close to active transcription start sites (TSSs) in the human genome. Transcription proceeds discontinuously with an accumulation of RNA polymerase II (RNAPII) in a paused state near the TSS. Consequently, replication forks inevitably encounter paused RNAPII soon after replication initiates. Hence, dedicated machinery may be needed to remove RNAPII and facilitate unperturbed fork progression. In this study, we discovered that Integrator, a transcription termination machinery involved in the processing of RNAPII transcripts, interacts with the replicative helicase at active forks and promotes the removal of RNAPII from the path of the replication fork. Integrator-deficient cells have impaired replication fork progression and accumulate hallmarks of genome instability including chromosome breaks and micronuclei. The Integrator complex resolves co-directional transcription-replication conflicts to facilitate faithful DNA replication.

AB - DNA replication preferentially initiates close to active transcription start sites (TSSs) in the human genome. Transcription proceeds discontinuously with an accumulation of RNA polymerase II (RNAPII) in a paused state near the TSS. Consequently, replication forks inevitably encounter paused RNAPII soon after replication initiates. Hence, dedicated machinery may be needed to remove RNAPII and facilitate unperturbed fork progression. In this study, we discovered that Integrator, a transcription termination machinery involved in the processing of RNAPII transcripts, interacts with the replicative helicase at active forks and promotes the removal of RNAPII from the path of the replication fork. Integrator-deficient cells have impaired replication fork progression and accumulate hallmarks of genome instability including chromosome breaks and micronuclei. The Integrator complex resolves co-directional transcription-replication conflicts to facilitate faithful DNA replication.

KW - fork reversal

KW - genome stability

KW - Integrator complex

KW - MCM complex

KW - R-loop

KW - replication stress

KW - replicative helicase

KW - RNA polymerase II

KW - RNAPII

KW - transcription-replication conflicts

KW - TRCs

U2 - 10.1016/j.molcel.2023.05.015

DO - 10.1016/j.molcel.2023.05.015

M3 - Journal article

C2 - 37295432

AN - SCOPUS:85163378344

SN - 1097-2765

VL - 83

SP - 2357-2366.e8

JO - Molecular Cell

JF - Molecular Cell

IS - 13

ER -