Đermić, Edyta; Zahradka, Davor; Vujaklija, Dušica; Ivanković, Siniša; Đermić, Damir (2017) 3'-terminated Overhangs Regulate DNA Double-Strand Break Processing in Escherichia coli. G3-Genes Genomes Genetics, 7 (9). pp. 3091-3102. ISSN 2160-1836
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Abstract
Double-strand breaks (DSBs) are lethal DNA lesions, which are repaired by homologous recombination in Escherichia coli To study DSB processing in vivo, we induced DSBs into the E. coli chromosome by gamma irradiation and measured chromosomal degradation. We show that the DNA degradation is regulated by RecA protein concentration and its rate of association with ssDNA. RecA decreased DNA degradation in wild- type, recB and recD strains, indicating that it is a general phenomenon in E. coli On the other hand, DNA degradation was greatly reduced and unaffected by RecA in the recB1080 mutant (which produces long overhangs) and in a strain devoid of four exonucleases that degrade a 3' tail (ssExos). 3'-5' ssExos deficiency is epistatic to RecA deficiency concerning DNA degradation, suggesting that bound RecA is shielding 3' tail from degradation by 3'-5' ssExos. Since 3'-tail preservation is common to all these situations, we infer that RecA polymerization constitutes a subset of mechanisms for preserving the integrity of 3' tails emanating from DSBs, along with 3' tail's massive length, or prevention of their degradation by inactivation of 3'-5' ssExos. Thus, we conclude that 3' overhangs are crucial in controlling the extent of DSB processing in E. coli This study suggests a regulatory mechanism for DSB processing in E. coli, wherein 3' tails impose a negative feedback loop on DSB processing reactions, specifically on helicase reloading onto dsDNA ends.
| Item Type: | Article | ||||||||
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| Uncontrolled Keywords: | DNA degradation ; exonuclease activity ; RecA protein ; single-strand specific exonucleases ; recB1080 mutant | ||||||||
| Subjects: | NATURAL SCIENCES > Biology NATURAL SCIENCES > Biology > Microbiology NATURAL SCIENCES > Biology > Genetics, Evolution and Phylogenetics |
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| Divisions: | Division of Molecular Biology Division of Molecular Medicine |
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| Depositing User: | Damir Đermić | ||||||||
| Date Deposited: | 09 Nov 2022 12:24 | ||||||||
| URI: | http://fulir.irb.hr/id/eprint/7611 | ||||||||
| DOI: | 10.1534/g3.117.043521 |
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