The Single-Stranded DNA Binding Protein of Sulfolobus solfataricus Acts in the Presynaptic Step of Homologous Recombination

Rolfsmeier, Michael; Haseltine, Cynthia A.

doi:10.1016/j.jmb.2010.01.004

Cited by 25 publications

(37 citation statements)

References 63 publications

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“…Archaeal recombinases and various HR-associated enzymes of archaea specifically resemble the eukaryotic, rather than bacterial, homologues (Brendel et al, 1997, Constantinesco et al, 2004. The ssDNAbinding proteins of euryarchaeotes resemble eukaryotic replication proteins A (RPA)s whereas those of crenarchaeotes resemble bacterial single-stranded binding proteins (SSBs) (Kerr et al, 2003;Rolfsmeier & Haseltine, 2010). Furthermore, genes for several HR proteins which can be deleted from bacteria and eukaryotes are essential in hyperthermophilic archaea (Fujikane et al, 2010;Zhang et al, 2013), suggesting that, in archaea, these proteins play unusually important roles.…”

Section: Introductionmentioning

confidence: 99%

Homologous recombination in the archaeon Sulfolobus acidocaldarius: effects of DNA substrates and mechanistic implications

2013

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Although homologous recombination (HR) is known to influence the structure, stability, and evolution of microbial genomes, few of its functional properties have been measured in cells of hyperthermophilic archaea. The present study manipulated various properties of the parental DNAs in high-resolution assays of Sulfolobus acidocaldarius transformation, and measured the impact on the efficiency and pattern of marker transfer to the recipient chromosome. The relative orientation of homologous sequences, the type and position of chromosomal mutation being replaced, and the length of DNA flanking the marked region all affected the efficiency, linkage, tract continuity, and other parameters of marker transfer. Effects predicted specifically by the classical reciprocal-exchange model of HR were not observed. One analysis observed only 90 % linkage between markers defined by adjacent bases; in another series of experiments, sequence divergence up to 4 % had no detectable impact on overall efficiency of HR or on the co-transfer of a distal non-selected marker. The effects of introducing DNA via conjugation, rather than transformation, were more difficult to assess, but appeared to increase co-transfer (i.e. linkage) of relatively distant non-selected markers. The results indicate that HR events between gene-sized duplex DNAs and the S. acidocaldarius chromosome typically involve neither crossing over nor interference from a mismatch-activated anti-recombination system. Instead, the donor DNA may anneal to a transient chromosomal gap, as in the mechanism proposed for oligonucleotidemediated transformation of Sulfolobus and other micro-organisms.

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Section: Introductionmentioning

confidence: 99%

Homologous recombination in the archaeon Sulfolobus acidocaldarius: effects of DNA substrates and mechanistic implications

2013

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“…23 The S. solfataricus RadA protein is a homologue of eukaryotic Rad51 protein and is involved in the HR-based repair of DNA DSBs. [31][32][33][34][35][36] The catalytic domain of S. solfataricus Rad54 protein has been crystallized and is structurally similar to that of the zebrafish Rad54 protein. 37,38 This protein is a double-strand DNA- .…”

Section: Resultsmentioning

confidence: 99%

Repair of DNA Double-Strand Breaks Induced by Ionizing Radiation Damage Correlates with Upregulation of Homologous Recombination Genes in Sulfolobus solfataricus

Rolfsmeier

Laughery

Haseltine

2011

Journal of Molecular Biology

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“…Several genes have been identified in Sulfolobus species that are expected to be involved in DSB repair in vivo. Biochemical studies have elucidated the activities of the RadA protein (a homologue of eukaryotic Rad51 and bacterial RecA proteins) from both S. solfataricus and Sulfolobus tokodaii and the Rad54 protein from S. solfataricus and suggest that these proteins are involved in the repair of DNA DSBs through homologous recombination (3,14,31,38,39,42,48). The Mre11 protein from Sulfolobus acidocaldarius has been found to be associated with DNA following damage and directly interacts with Rad50, implying a role in break repair for both of these proteins (28).…”

Section: Resultsmentioning

confidence: 99%

Repair of DNA Double-Strand Breaks following UV Damage in ThreeSulfolobussolfataricusStrains

2010

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DNA damage repair mechanisms have been most thoroughly explored in the eubacterial and eukaryotic branches of life. The methods by which members of the archaeal branch repair DNA are significantly less well understood but have been gaining increasing attention. In particular, the approaches employed by hyperthermophilic archaea have been a general source of interest, since these organisms thrive under conditions that likely lead to constant chromosomal damage. In this work we have characterized the responses of three Sulfolobus solfataricus strains to UV-C irradiation, which often results in double-strand break formation. We examined S. solfataricus strain P2 obtained from two different sources and S. solfataricus strain 98/2, a popular strain for site-directed mutation by homologous recombination. Cellular recovery, as determined by survival curves and the ability to return to growth after irradiation, was found to be strain specific and differed depending on the dose applied. Chromosomal damage was directly visualized using pulsed-field gel electrophoresis and demonstrated repair rate variations among the strains following UV-C irradiation-induced double-strand breaks. Several genes involved in double-strand break repair were found to be significantly upregulated after UV-C irradiation. Transcript abundance levels and temporal expression patterns for doublestrand break repair genes were also distinct for each strain, indicating that these Sulfolobus solfataricus strains have differential responses to UV-C-induced DNA double-strand break damage.Cells have evolved molecular mechanisms to meet the challenge of maintaining genomic integrity by rapidly responding to environmental stresses that can damage proteins and DNA. One of the most common forms of damage is caused by UV light (UV) exposure. High-energy short-wavelength UV-C light is absorbed directly by DNA and induces both cyclobutane pyrimidine dimers between adjacent thymidine or cytosine residues as well as pyrimidine-pyrimidone photoproducts between adjacent pyrimidine residues. Mechanisms for repair of these lesions appear to be present in all organisms and are thought to occur through either light-independent nucleotide excision repair (NER) or light-dependent photoreactivation using photolyases (for reviews, see references 33 and 34). UV-C irradiation also causes the production of reactive oxygen species, which can result in DNA double-strand breaks (DSBs) (6, 44). Our primary understanding for repair of DSBs has come from studies focused primarily on bacteria and eukaryotes. In Escherichia coli, these breaks are repaired through the action of the RecA protein, which assists in recombinational repair of single-strand regions produced through replication fork arrest at UV lesions and in DSB repair by extended synthesis-dependent strand annealing (SDSA) and homologous recombination (9, 19). Eukaryotes employ nonhomologous end joining as well as DSB repair by SDSA and homologous recombination mechanisms to repair these breaks (for recent reviews, see ref...

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The Single-Stranded DNA Binding Protein of Sulfolobus solfataricus Acts in the Presynaptic Step of Homologous Recombination

Cited by 25 publications

References 63 publications

Homologous recombination in the archaeon Sulfolobus acidocaldarius: effects of DNA substrates and mechanistic implications

Homologous recombination in the archaeon Sulfolobus acidocaldarius: effects of DNA substrates and mechanistic implications

Repair of DNA Double-Strand Breaks Induced by Ionizing Radiation Damage Correlates with Upregulation of Homologous Recombination Genes in Sulfolobus solfataricus

Repair of DNA Double-Strand Breaks following UV Damage in ThreeSulfolobussolfataricusStrains

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