Characteristics of Purified recA Protein and the Regulation of Its Synthesis In Vivo

Ogawa, Tomoko; Wabiko, Hiroetsu; Tsurimoto, Toshiki; Horii, Toshihiro; Masukata, Hisao; Ogawa, Haruko

doi:10.1101/sqb.1979.043.01.099

Cited by 192 publications

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“…Strand exchange is associated with the unidirectional extension of the joint (unidirectional branch migration) by several kilobase pairs (34,35) and can integrate mismatches (36) and heterologous sequences (37) into a heteroduplex. RecA has DNA-dependent ATPase activity (38), and ATP is required for heteroduplex joint formation (11,12,35). ATP hydrolysis is not required for homologous pairing but is required for unidirectional branch migration to bypass heterologous sequences (35,39,40), and ATP hydrolysis stimulates the detachment of the protein from the DNA for the recycling of RecA (41)(42)(43).…”

Section: Heteroduplex Joint Formation Is Promoted By the Reca͞rad51-fmentioning

confidence: 99%

Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: A possible advantage of DNA over RNA as genomic material

Shibata

Nishinaka

Mikawa

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Heteroduplex joints are general intermediates of homologous genetic recombination in DNA genomes. A heteroduplex joint is formed between a single-stranded region (or tail), derived from a cleaved parental double-stranded DNA, and homologous regions in another parental double-stranded DNA, in a reaction mediated by the RecA͞Rad51-family of proteins. In this reaction, a RecA͞ Rad51-family protein first forms a filamentous complex with the single-stranded DNA, and then interacts with the double-stranded DNA in a search for homology. Studies of the three-dimensional structures of single-stranded DNA bound either to Escherichia coli RecA or Saccharomyces cerevisiae Rad51 have revealed a novel extended DNA structure. This structure contains a hydrophobic interaction between the 2 methylene moiety of each deoxyribose and the aromatic ring of the following base, which allows bases to rotate horizontally through the interconversion of sugar puckers. This base rotation explains the mechanism of the homology search and base-pair switch between double-stranded and singlestranded DNA during the formation of heteroduplex joints. The pivotal role of the 2 methylene-base interaction in the heteroduplex joint formation is supported by comparing the recombination of RNA genomes with that of DNA genomes. Some simple organisms with DNA genomes induce homologous recombination when they encounter conditions that are unfavorable for their survival. The extended DNA structure confers a dynamic property on the otherwise chemically and genetically stable double-stranded DNA, enabling gene segment rearrangements without disturbing the coding frame (i.e., protein-segment shuffling). These properties may give an extensive evolutionary advantage to DNA.base-pair switch ͉ homologous pairing ͉ strand exchange ͉ NMR ͉ three-dimensional molecular structure DNA as a General Molecular Carrier of Genetic Information G enomic information generally is carried by double-stranded DNA. The double-stranded DNA structure discovered by Watson and Crick clearly explains the mechanisms of heredity, which include both the encoding of genetic information and its duplication as chemical properties of the molecule (1). In addition, the double-stranded structure enables cellular systems to recognize, as structural irregularities, erroneously incorporated nucleotides or lesions in bases, sugars, or the backbone strand, and to correct these errors by using the partner strand as a template. The genetic stability and the chemical inactivity of double-stranded DNA have been regarded as favorable molecular properties for its role as the carrier of genomic information. Evolution, which is a general attribute of the genome as well, has resulted in a variety of organisms, whose diversity arose not only as a result of changes in the genomic information, but also as a result of increased content and complexity. The faithful duplication and repair exhibited by the double-stranded DNA structure would seem to be incompatible with the process of evolution. Thus, evolution has b...

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Section: Heteroduplex Joint Formation Is Promoted By the Reca͞rad51-fmentioning

confidence: 99%

Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: A possible advantage of DNA over RNA as genomic material

Shibata

Nishinaka

Mikawa

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…Furthermore, resistance to UV-induced damage of this mutant could not be restored to wild-type levels by complementation with a functional copy of recA from plasmid pCT303, even though introduction of pCT303 into E. coli DK1 (which has a deletion of the recA gene) completely restored UV resistance. Previous in vitro work has shown that the singlestranded DNA-dependent ATPase activity of RecA is a property of the tetrameric form of the protein (Ogawa et al, 1978). Thus ATPase activity is required for the in vitro recombination functions of purified RecA (Radding, 1982 ;Weinstock et al, 1979), although this is not the case for the proteolytic activity of RecA involved in SOS induction (Phizicky & Roberts, 1981).…”

Section: Discussionmentioning

confidence: 79%

“…ATP binding is also required for formation of RecA tetramers and filamentous aggregates of the RecA protein (Ogawa et al, 1978). When RecA1 missense protein and wildtype RecA protein of E. coli form mixed tetramers in the Analysis of recA from X. bovienii presence of ATP, a dramatic reduction in ATPase activity is observed (Ogawa et al, 1978). Phenotypically, the recA1 mutation, Gly-160 to Asp-160, is indistinguishable from a recA deletion (Kowalczykowski et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

“…Thus ATPase activity is required for the in vitro recombination functions of purified RecA (Radding, 1982 ;Weinstock et al, 1979), although this is not the case for the proteolytic activity of RecA involved in SOS induction (Phizicky & Roberts, 1981). ATP binding is also required for formation of RecA tetramers and filamentous aggregates of the RecA protein (Ogawa et al, 1978). When RecA1 missense protein and wildtype RecA protein of E. coli form mixed tetramers in the Analysis of recA from X. bovienii presence of ATP, a dramatic reduction in ATPase activity is observed (Ogawa et al, 1978).…”

Section: Discussionmentioning

confidence: 99%

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Xenorhabdus bovienii T228 phase variation and virulence are independent of RecA function

2000

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5005Colony pleomorphism, or phase variation, expressed by entomopathogenic bacteria belonging to the genus Xenorhabdus, is an important factor which determines the association of the bacteria with their nematode symbiont and the outcome of infection of susceptible insect larvae by the bacteriumnematode parasitic complex. The mechanism underlying phase variation is unknown. To determine whether RecA-mediated processes are linked to phase variation, the recA gene of Xenorhabdus bovienii was cloned and sequenced. When expressed in a recA-deleted strain of Escherichia coli, the X. bovienii recA clone was able to complement the loss of RecA function. X. bovienii chromosomal recA insertion mutants showed increased sensitivity to UV. Phase 1 forms did not show altered ability to convert to phase 2 and no significant differences in expression of other phase-dependent characteristics, including phospholipase C, haemolysin, protease, antibiotic activity and Congo Red binding, were noted. Furthermore, the LD 50 of the X. bovienii recA insertion mutant for Galleria mellonella larvae was not significantly different from that of wild-type strains. From these data the authors conclude that recA is unlikely to be involved in phase variation, the expression of phasedependent characteristics, or virulence factors involved in killing of susceptible larvae.

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“…Mutations in RecA were first identified in 1969 (152) and the basic RecA strand exchange activity was first described in 1979 (93,101,119). Mutations in RecA's eukaryotic homolog, Rad51, were found in budding yeast in 1974 (29), but it was not until 1992 that yeast Rad51 was definitively identified as the eukaryotic homolog of RecA and not until 1994 that its biochemical activity was established (1, 134).…”

Section: Introductionmentioning

confidence: 99%

A Life Investigating Pathways That Repair Broken Chromosomes

Haber

2016

Annu. Rev. Genet.

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Double-strand breaks (DSBs) pose a severe challenge to genome integrity; consequently, cells have developed efficient mechanisms to repair DSBs through several pathways of homologous recombination and other nonhomologous end-joining processes. Much of our understanding of these pathways has come from the analysis of site-specific DSBs created by the HO endonuclease in the budding yeast Saccharomyces cerevisiae. I was fortunate to get in on the ground floor of analyzing the fate of synchronously induced DSBs through the study of what I coined "in vivo biochemistry." I have had the remarkable good fortune to profit from the development of new techniques that have permitted an ever more detailed dissection of these repair mechanisms, which are described here.

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Characteristics of Purified recA Protein and the Regulation of Its Synthesis In Vivo

Cited by 192 publications

References 0 publications

Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: A possible advantage of DNA over RNA as genomic material

Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: A possible advantage of DNA over RNA as genomic material

Xenorhabdus bovienii T228 phase variation and virulence are independent of RecA function

A Life Investigating Pathways That Repair Broken Chromosomes

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