Format

Send to

Choose Destination
EMBO Rep. 2015 Feb;16(2):221-31. doi: 10.15252/embr.201439458. Epub 2014 Dec 19.

RNA-processing proteins regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA.

Author information

1
Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy.
2
Institut Curie, CNRS UMR3244 Université Pierre et Marie Curie, Paris Cedex 05, France.
3
Institut Curie, CNRS UMR3244 Université Pierre et Marie Curie, Paris Cedex 05, France antonin.morillon@curie.fr fabrizio.dadda@ifom.eu mariapia.longhese@unimib.it.
4
IFOM Foundation-FIRC Institute of Molecular Oncology Foundation, Milan, Italy Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Pavia, Italy antonin.morillon@curie.fr fabrizio.dadda@ifom.eu mariapia.longhese@unimib.it.
5
Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy antonin.morillon@curie.fr fabrizio.dadda@ifom.eu mariapia.longhese@unimib.it.

Abstract

Eukaryotic cells respond to DNA double-strand breaks (DSBs) by activating a checkpoint that depends on the protein kinases Tel1/ATM and Mec1/ATR. Mec1/ATR is activated by RPA-coated single-stranded DNA (ssDNA), which arises upon nucleolytic degradation (resection) of the DSB. Emerging evidences indicate that RNA-processing factors play critical, yet poorly understood, roles in genomic stability. Here, we provide evidence that the Saccharomyces cerevisiae RNA decay factors Xrn1, Rrp6 and Trf4 regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA. The lack of Xrn1 inhibits ssDNA generation at the DSB by preventing the loading of the MRX complex. By contrast, DSB resection is not affected in the absence of Rrp6 or Trf4, but their lack impairs the recruitment of RPA, and therefore of Mec1, to the DSB. Rrp6 and Trf4 inactivation affects neither Rad51/Rad52 association nor DSB repair by homologous recombination (HR), suggesting that full Mec1 activation requires higher amount of RPA-coated ssDNA than HR-mediated repair. Noteworthy, deep transcriptome analyses do not identify common misregulated gene expression that could explain the observed phenotypes. Our results provide a novel link between RNA processing and genome stability.

KEYWORDS:

DNA damage checkpoint; DNA double‐strand breaks; Rrp6; Trf4; Xrn1

PMID:
25527408
PMCID:
PMC4328749
DOI:
10.15252/embr.201439458
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center