Involvement of ATM in homologous recombination after end resection and RAD51 nucleofilament formation.

Abstract

Ataxia-telangiectasia mutated (ATM) is needed for the initiation of the double-strand break (DSB) repair by homologous recombination (HR). ATM triggers DSB end resection by stimulating the nucleolytic activity of CtIP and MRE11 to generate 3'-ssDNA overhangs, followed by RPA loading and RAD51 nucleofilament formation. Here we show for the first time that ATM is also needed for later steps in HR after RAD51 nucleofilament formation. Inhibition of ATM after completion of end resection did not affect RAD51 nucleofilament formation, but resulted in HR deficiency as evidenced by (i) an increase in the number of residual RAD51/γH2AX foci in both S and G2 cells, (ii) the decrease in HR efficiency as detected by HR repair substrate (pGC), (iii) a reduced SCE rate and (iv) the radiosensitization of cells by PARP inhibition. This newly described role for ATM was found to be dispensable in heterochromatin-associated DSB repair, as KAP1-depletion did not alleviate the HR-deficiency when ATM was inhibited after end resection. Moreover, we demonstrated that ATR can partly compensate for the deficiency in early, but not in later, steps of HR upon ATM inhibition. Taken together, we describe here for the first time that ATM is needed not only for the initiation but also for the completion of HR.

• Authors: A Bakr, C Oing, S Köcher, K Borgmann, I Dornreiter, C Petersen, E Dikomey, W Y Mansour
• Journal: Nucleic acids research (Nucleic Acids Res) Vol. 43 Issue 6 Pg. 3154-66 (Mar 31 2015) ISSN: 1362-4962 [Electronic] England
• PMID: 25753674 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Chemical References

• 2-morpholin-4-yl-6-thianthren-1-yl-pyran-4-one
• 6-(4-hydroxyphenyl)-2-thioxo-2,3-dihydro-4(1H)-pyrimidinone
• DNA-Binding Proteins
• Heterochromatin
• MRE11 protein, human
• Morpholines
• Pyrimidinones
• Pyrones
• Thiones
• ATM protein, human
• Ataxia Telangiectasia Mutated Proteins
• RAD51 protein, human
• Rad51 Recombinase
• MRE11 Homologue Protein

Topics

• Ataxia Telangiectasia Mutated Proteins (antagonists & inhibitors, genetics, metabolism)
• Cell Line
• DNA Breaks, Double-Stranded
• DNA Repair
• DNA-Binding Proteins (antagonists & inhibitors, genetics, metabolism)
• Epistasis, Genetic
• G2 Phase
• Gene Knockdown Techniques
• HeLa Cells
• Heterochromatin (genetics, metabolism)
• Homologous Recombination (drug effects)
• Humans
• MRE11 Homologue Protein
• Morpholines (pharmacology)
• Pyrimidinones (pharmacology)
• Pyrones (pharmacology)
• Rad51 Recombinase (metabolism)
• S Phase
• Thiones (pharmacology)