caa a22 4500 445818832 CHVBK 20180317145237.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s00792-010-0351-2 doi (NATIONALLICENCE)springer-10.1007/s00792-010-0351-2 The carboxyl terminal of the archaeal nuclease NurA is involved in the interaction with single-stranded DNA-binding protein and dimer formation [Elektronische Daten] [Tao Wei, Songtao Zhang, Linlin Hou, Jinfeng Ni, Duohong Sheng, Yulong Shen] The nuclease NurA is present in all known thermophilic archaea and has been implicated to facilitate efficient DNA double-strand break end processing in Mre11/Rad50-mediated homologous recombinational repair. To understand the structural and functional relationship of this enzyme, we constructed five site-directed mutants of NurA from Sulfolobus tokodaii (StoNurA), D56A, E114A, D131A, Y291A, and H299A, at the conserved motifs, and four terminal deletion mutants, StoNurAΔN (19-331), StoNurAΔNΔC (19-303), StoNurAΔC (1-281), and StoNurAΔC (1-303), and characterized the proteins biochemically. We found that mutation at the acidic residue, D56, E114, D131, or at the basic residue, H299, abolishes the nuclease activity, while mutation at the aromatic residue Y291 only impairs the activity. Interestingly, by chemical cross-linking assay, we found that the mutant Y291A is unable to form stable dimer. Additionally, we demonstrated that deletion of the C-terminal amino acid residues 304-331 of StoNurA results in loss of the physical and functional interaction with the single-stranded DNA-binding protein (StoSSB). These results established that the C-terminal conserved aromatic residue Y291 is involved in dimer formation and the C-terminal residues 304-331 of NurA are involved in the interaction with single-stranded DNA-binding protein. Springer, 2010 StoNurA nationallicence StoSSB nationallicence Homologous recombination nationallicence DNA repair nationallicence Sulfolobus tokodaii nationallicence Wei Tao State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Zhang Songtao State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Hou Linlin State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Ni Jinfeng State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Sheng Duohong State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Shen Yulong State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut Extremophiles Springer Japan 15/2(2011-03-01), 227-234 1431-0651 15:2<227 2011 15 792 https://doi.org/10.1007/s00792-010-0351-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00792-010-0351-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wei Tao State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Songtao State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 700 1- Hou Linlin State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 700 1- Ni Jinfeng State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 700 1- Sheng Duohong State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 700 1- Shen Yulong State Key Laboratory of Microbial Technology, Shandong University, 27 Shanda Nan Road, 250100, Jinan, People's Republic of China aut NATIONALLICENCE 773 0- Extremophiles Springer Japan 15/2(2011-03-01), 227-234 1431-0651 15:2<227 2011 15 792 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer