Physical and functional interaction between archaeal single-stranded DNA-binding protein and the 5'-3' nuclease NurA.
NurA is a novel 5'-3' exonuclease that is closely linked to Mre11 and Rad50 homologues in most thermophilic archaea. We report a physical and functional interaction between NurA (StoNurA) and single-stranded DNA-binding protein (StoSSB) from the hyperthermophilic archaeon Sulfolobus tokodaii. StoSSB was identified as a novel StoNurA-interacting protein by pull-down assay using Ni-NTA agarose beads and MALDI-TOF mass spectrometry. The direct interaction between StoNurA and StoSSB was further confirmed by yeast two-hybrid and co-immunoprecipitation analysis. The interaction was supposed to have functional significance because it was found that StoSSB inhibited the 5'-3' ssDNA and dsDNA exonuclease and ssDNA endonuclease activities of StoNurA. Our results suggest that NurA may function closely together with SSB in DNA transactions in archaea.
Wei
Tao
T
State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, Jinan, 250100, PR China.
Zhang
Songtao
S
Zhu
Shanshan
S
Sheng
Duohong
D
Ni
Jinfeng
J
Shen
Yulong
Y
D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D005090:Exodeoxyribonucleases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D047468:Immunoprecipitation; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D016944:Sulfolobus / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D019843:Archaeal Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D004247:DNA; D005090:Exodeoxyribonucleases