title: Biochemical characterization of RNase HII from Aeropyrum pernix.
abstract: Aeropyrum pernix contains one homolog of ribonuclease H (RNase H), A. pernix RNase HII (Ape-RNase HII). Activity characterization showed that Ape-RNase HII exhibited the highest activity in the presence of 5 mM Mn(2+), 1 mM Co(2+), or 10 mM Mg(2+), respectively; however, its cleavage efficiencies at different cleavage sites for Mn(2+) and Mg(2+) were different. Ape-RNase HII cleaved 12-bp RNA/DNA substrates at multiple sites and the optimum pH value was 11.0. Moreover, 16-bp DNA-r4-DNA/DNA and 13-bp DNA-r1-DNA/DNA chimeric substrates were cleaved at DNA-RNA junction. Ape-RNase HII was thermostable and the stabilization was enhanced with increased salt concentration. This work is believed to be the first in vitro functional study of Ape-RNase HII and the results should contribute to the analysis of RNase H of other archaeal species.
authors.affiliation: Instrumental Analysis Center of Shanghai Jiaotong University, Shanghai, China.
authors.forename: Jingli
authors.initials: J
authors.lastname: Hou
authors.forename: Yufen
authors.initials: Y
authors.lastname: Liu
authors.forename: Zheng
authors.initials: Z
authors.lastname: Lu
authors.forename: Xipeng
authors.initials: X
authors.lastname: Liu
authors.forename: Jianhua
authors.initials: J
authors.lastname: Liu
mesh_terms: D041522:Aeropyrum / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D055162:Biocatalysis / Q000187:drug effects; D003035:Cobalt / Q000494:pharmacology; D019641:DNA, Archaeal / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology; D008969:Molecular Sequence Data; D019642:RNA, Archaeal / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016914:Ribonuclease H / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D019642:RNA, Archaeal; D011994:Recombinant Proteins; D003035:Cobalt; D008345:Manganese; C091139:ribonuclease HII; D016914:Ribonuclease H