Hypoosmotic stress induces R loop formation in nucleoli and ATR/ATM-dependent silencing of nucleolar transcription

Artem K Velichko; Nadezhda V Petrova; Artem V Luzhin; Olga S Strelkova; Natalia Ovsyannikova; Igor I Kireev; Natalia V Petrova; Sergey V Razin; Omar L Kantidze

doi:10.1093/nar/gkz436

Hypoosmotic stress induces R loop formation in nucleoli and ATR/ATM-dependent silencing of nucleolar transcription

Nucleic Acids Res. 2019 Jul 26;47(13):6811-6825. doi: 10.1093/nar/gkz436.

Authors

Artem K Velichko^{1

2}, Nadezhda V Petrova¹, Artem V Luzhin¹, Olga S Strelkova³, Natalia Ovsyannikova³, Igor I Kireev^{3

4}, Natalia V Petrova⁵, Sergey V Razin^{5

6

7}, Omar L Kantidze^{1

7}

Affiliations

¹ Laboratory of Genome Stability, Institute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, Russia.
² Institute for Translational Medicine and Biotechnology, Sechenov First Moscow State Medical University, 119991 Moscow, Russia.
³ A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119992 Moscow, Russia.
⁴ V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology, and Perinatology, 117997 Moscow, Russia.
⁵ Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, Russia.
⁶ Department of Molecular Biology, Moscow State University, 119234 Moscow, Russia.
⁷ LFR2O, Institute Gustave Roussy, F-94805 Villejuif, France.

Abstract

The contribution of nucleoli to the cellular stress response has been discussed for over a decade. Stress-induced inhibition of RNA polymerase I-dependent transcription is hypothesized as a possible effector program in such a response. In this study, we report a new mechanism by which ribosomal DNA transcription can be inhibited in response to cellular stress. Specifically, we demonstrate that mild hypoosmotic stress induces stabilization of R loops in ribosomal genes and thus provokes the nucleoli-specific DNA damage response, which is governed by the ATM- and Rad3-related (ATR) kinase. Activation of ATR in nucleoli strongly depends on Treacle, which is needed for efficient recruitment/retention of TopBP1 in nucleoli. Subsequent ATR-mediated activation of ATM results in repression of nucleolar transcription.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Ataxia Telangiectasia Mutated Proteins / physiology*
Carrier Proteins / genetics*
Cell Line
Cell Nucleolus / drug effects
Cell Nucleolus / metabolism*
Cell Survival
DNA Breaks, Double-Stranded
DNA Damage
DNA Replication
DNA, Ribosomal / genetics*
DNA-Binding Proteins / genetics*
Dactinomycin / pharmacology
Enzyme Activation / drug effects
Gene Knockout Techniques
Gene Silencing*
Histones / metabolism
Humans
Hypotonic Solutions / pharmacology
Mice
Nuclear Proteins / genetics*
Nuclear Proteins / physiology
Osmotic Pressure*
Phosphoproteins / physiology
Phosphorylation / drug effects
Protein Kinase Inhibitors / pharmacology
Protein Processing, Post-Translational / drug effects
R-Loop Structures*
Transcription, Genetic / drug effects
Transcription, Genetic / physiology*

Substances

Carrier Proteins
DNA, Ribosomal
DNA-Binding Proteins
H2AX protein, human
Histones
Hypotonic Solutions
Nuclear Proteins
Phosphoproteins
Protein Kinase Inhibitors
TCOF1 protein, human
TOPBP1 protein, human
Dactinomycin
ATM protein, human
ATR protein, human
Ataxia Telangiectasia Mutated Proteins