Archaeal DNA helicase HerA interacts with Mre11 homologue and unwinds blunt-ended double-stranded DNA and recombination intermediates

Zhang, Songtao; Wei, Tao; Hou, Guihua; Zhang, Chao; Liang, Pengjuan; Ni, Jinfeng; Sheng, Duohong; Shen, Yulong

doi:10.1016/j.dnarep.2007.10.010

Cited by 30 publications

(40 citation statements)

References 31 publications

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“…HerA is an ATP-dependent bipolar helicase in hyperthermophilic archaea (16,17,55). We found that DrHerA has ATPase activity, based on free-P i release measurements.…”

Section: Resultsmentioning

confidence: 99%

“…Given that mesophilic features were seen for DrHerA ATPase activity, we also performed nuclease assays at 30,35,40,45,50,55, and 60°C; the optimum reaction temperature was around 55°C (Fig. 5C).…”

Section: Resultsmentioning

confidence: 99%

“…Because both DrHerA and DrNurA showed mesophilic features in vitro, we studied the effects for HerA and NurA mutants growing at a normal culturing temperature (30°C) and a higher culturing temperature (37°C). nurA, herA, and the nurA-herA operon were knocked out by deletion replacement and verified by PCR (see (30,35,40,45,50,55, and 60°C) were tested. (Left) The different nuclease activities were compared at different temperatures by using ssDNA as the substrate.…”

Section: Resultsmentioning

confidence: 99%

“…An ATPase assay was performed as previously described (45), which was measured by spectrophotometric detection of inorganic phosphate (P i ) with malachite green reagent 35,40,45,50,55, and 60°C were used.) Malachite green reagent (200 ml in 700 ml water) was added, and P i release was monitored at 630 nm.…”

Section: Methodsmentioning

confidence: 99%

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Biochemical and Functional Characterization of the NurA-HerA Complex from Deinococcus radiodurans

Cheng

Chen

et al. 2015

J Bacteriol

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In archaea, the NurA nuclease and HerA ATPase/helicase, together with the Mre11-Rad50 complex, function in 3= singlestranded DNA (ssDNA) end processing during homologous recombination (HR). However, bacterial homologs of NurA and HerA have not been characterized. From Deinococcus radiodurans, we identified the manganese-dependent 5=-to-3= ssDNA/double-stranded DNA (dsDNA) exonuclease/endonuclease NurA (DrNurA) and the ATPase HerA (DrHerA). These two proteins stimulated each other's activity through direct protein-protein interactions. The N-terminal HAS domain of DrHerA was the key domain for this interaction. Several critical residues of DrNurA and DrHerA were verified by site-directed mutational analysis. Temperature-dependent activity assays confirmed that the two proteins had mesophilic features, with optimum activity temperatures 10°C to 15°C higher than their optimum growth temperatures. Knocking out either nurA or herA affected cell proliferation by shortening the growth phase, especially for growth at a high temperature (37°C). In addition, both mutant strains displayed almost 10-fold-reduced intermolecular recombination efficiency, indicating that DrNurA and DrHerA might be involved in homologous recombination in vivo. However, single-and double-gene deletions did not show significantly decreased radioresistance. Our results confirmed that the biochemical activities of bacterial NurA and HerA proteins were conserved with archaea. Our phenotypical results suggested that these proteins might have different functions in bacteria. IMPORTANCEDeinococcus radiodurans NurA (DrNurA) was identified as a manganese-dependent 5=-to-3= ssDNA/dsDNA exonuclease/endonuclease, and Deinococcus radiodurans HerA (DrHerA) was identified as an ATPase. Physical interactions between DrNurA and DrHerA explained mutual stimulation of their activities. The N-terminal HAS domain on DrHerA was identified as the interaction domain. Several essential functional sites on DrNurA and DrHerA were characterized. Both DrHerA and DrNurA showed mesophilic biochemical features, with their optimum activity temperatures 10°C to 15°C higher than their optimum growth temperatures in vitro. Knockout of nurA or herA led to abnormal cell proliferation and reduced intermolecular recombination efficiency but no obvious effect on radioresistence.

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“…HerA is an ATP-dependent bipolar helicase in hyperthermophilic archaea (16,17,55). We found that DrHerA has ATPase activity, based on free-P i release measurements.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Biochemical and Functional Characterization of the NurA-HerA Complex from Deinococcus radiodurans

Cheng

Chen

et al. 2015

J Bacteriol

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“…The 84 mer ssDNA, 84 bp double stranded (ds) DNA, and four 72 mer oligonucleotides, which could form Holliday junction DNA by annealing, were consistent with the DNA substrates used in our previous publication [17]. The DNA substrates used here were -32 P-labeled at their 5′ ends as described previously [18], and stored in TE buffer (10 mmol L 1 Tris-HCl, pH 7.0, and 0.5 mmol L 1 EDTA).…”

Section: Dna Substratesmentioning

confidence: 49%

Sulfolobus tokodaii RadA paralog, stRadC2, is involved in DNA recombination via interaction with RadA and Hjc

Wang

Sheng

Han

et al. 2012

Sci. China Life Sci.

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Rad51/RadA paralogs found in eukaryotes and euryarchaea play important roles during recombination and repair, and mutations in one of the human Rad51 paralogs, Rad51C, are associated with breast and ovarian cancers. The hyperthermophilic crenarchaeon Sulfolobus tokodaii encodes four putative RadA paralogs and studies on these proteins may assist in understanding the functions of human Rad51 paralogs. Here, we report the biochemical characterization of stRadC2, a S. tokodaii RadA paralog. Pull-down assays revealed that the protein was able to interact with the recombinase, RadA, and the Holliday junction endonuclease, Hjc. stRadC2 inhibited the strand exchange activity of RadA and facilitated Hjc-mediated Holliday junction DNA cleavage in vitro. RT-PCR analysis revealed that stRadC2 transcription was immediately reduced after UV irradiation, but was restored to normal levels at the late stages of DNA repair. Our results suggest that stRadC2 may act as an anti-recombination factor in DNA recombinational repair in S. tokodaii.

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The Family Sulfolobaceae

Albers¹,

Siebers²

2014

The Prokaryotes

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Archaeal DNA helicase HerA interacts with Mre11 homologue and unwinds blunt-ended double-stranded DNA and recombination intermediates

Cited by 30 publications

References 31 publications

Biochemical and Functional Characterization of the NurA-HerA Complex from Deinococcus radiodurans

Biochemical and Functional Characterization of the NurA-HerA Complex from Deinococcus radiodurans

Sulfolobus tokodaii RadA paralog, stRadC2, is involved in DNA recombination via interaction with RadA and Hjc

The Family Sulfolobaceae

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