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In Escherichia coli, three isoforms of the essential translation initiation factor IF2 (IF2-1, IF2-2, and IF2-3) are generated from separate in-frame initiation codons in infB . The isoforms have earlier been suggested to additionally participate in DNA damage repair and replication restart. It is also known that the proteins RecA and RecBCD are needed for repair of DNA double-strand breaks (DSBs) in E. coli. Here, we show that strains lacking IF2-1 are profoundly sensitive to two-ended DSBs in DNA generated by radiomimetic agents phleomycin or bleomycin, or by endonuclease I-SceI. However, these strains remained tolerant to other DSB-generating genotoxic agents or perturbations to which recA and recBC mutants remained sensitive, such as to mitomycin C, type-2 DNA topoisomerase inhibitors, or DSB caused by palindrome cleavage behind a replication fork. Data from genome-wide copy number analyses following I-SceI cleavage at a single chromosomal locus suggested that, in a strain lacking IF2-1, the magnitude of recombination-dependent replication through replication restart mechanisms is largely preserved but the extent of DNA resection around the DSB site is reduced. We propose that in the absence of IF2-1 it is the synapsis of a RecA nucleoprotein filament to its homologous target that is weakened, which in turn leads to a specific failure in assembly of Ter -to- oriC directed replisomes needed for consummation of two-ended DSB repair. IMPORTANCE Double-strand breaks (DSBs) in DNA are major threats to genome integrity. In Escherichia coli, DSBs are repaired by RecA- and RecBCD-mediated homologous recombination (HR). This study demonstrates a critical role for an isoform (IF2-1) of the translation initiation factor IF2 in the repair of two-ended DSBs in E. coli (that can be generated by ionizing radiation, certain DNA-damaging chemicals, or endonuclease action). It is proposed that IF2-1 acts to facilitate the function of RecA in the synapsis between a pair of DNA molecules during HR.

Titel:	Essential Role for an Isoform of Escherichia coli Translation Initiation Factor IF2 in Repair of Two-Ended DNA Double-Strand Breaks.
Autor/in / Beteiligte Person:	Mallikarjun, J ; Gowrishankar, J
Zeitschrift:	Journal of bacteriology, Jg. 204 (2022-04-19), Heft 4, S. e0057121
Veröffentlichung:	Washington, DC : American Society for Microbiology, 2022
Medientyp:	academicJournal
ISSN:	1098-5530 (electronic)
DOI:	10.1128/jb.00571-21
Schlagwort:	DNA metabolism DNA Repair DNA Replication Peptide Initiation Factors genetics Peptide Initiation Factors metabolism Protein Isoforms genetics Protein Isoforms metabolism DNA Breaks, Double-Stranded Escherichia coli genetics Escherichia coli metabolism
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, Non-U.S. Gov't Language: English [J Bacteriol] 2022 Apr 19; Vol. 204 (4), pp. e0057121. <i>Date of Electronic Publication: </i>2022 Mar 28. MeSH Terms: DNA Breaks, Double-Stranded* ; Escherichia coli* / genetics ; Escherichia coli* / metabolism ; DNA / metabolism ; DNA Repair ; DNA Replication ; Peptide Initiation Factors / genetics ; Peptide Initiation Factors / metabolism ; Protein Isoforms / genetics ; Protein Isoforms / metabolism References: J Biol Chem. 2006 Jun 2;281(22):15238-48. (PMID: 16597620) ; Cell. 2011 Aug 19;146(4):533-43. (PMID: 21854980) ; Mol Microbiol. 1995 May;16(3):373-84. (PMID: 7565099) ; Biomed Res Int. 2014;2014:195162. (PMID: 25136561) ; J Mol Biol. 1975 Jan 5;91(1):53-66. (PMID: 1102696) ; Microbiol Mol Biol Rev. 2008 Dec;72(4):642-71, Table of Contents. (PMID: 19052323) ; EMBO J. 2021 May 17;40(10):e104847. 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(PMID: 7618864) Grant Information: DBT Centre of Excellence (COE) project for Microbial Biology; CRG/2018/000348 SERB project; BT/PR34340/BRB/10/1815/2019 Department of Biotechnology, Ministry of Science and Technology Contributed Indexing: Keywords: DNA repair; double-strand breaks; translation initiation factor IF2 Substance Nomenclature: 0 (Peptide Initiation Factors) ; 0 (Protein Isoforms) ; 9007-49-2 (DNA) Entry Date(s): Date Created: 20220328 Date Completed: 20220421 Latest Revision: 20220929 Update Code: 20231215 PubMed Central ID: PMC9017324

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Essential Role for an Isoform of Escherichia coli Translation Initiation Factor IF2 in Repair of Two-Ended DNA Double-Strand Breaks.