,Biochemical characterization of RNase HⅡ from Aeropyrum pernix

• 作 者: Jingli Hou Yufen Liu Zheng Lu Xipeng Liu Jianhua Liu
• 机 构: Instrumental Analysis Center of Shanghai Jiaotong University, Shanghai 200240, China“,”College of Li
• 出 处: 生物化学与生物物理学报（英文版）
• 发表日期: 2012年4期
• 关键词: RNase HⅡ Aeropyrum pernix thermostable RNA/DNA hybrid

Aeropyrum peix contains one homolog of ribonuclease H (RNase H),A.peix RNase HⅡ (Ape-RNase HⅡ).Activity characterization showed that Ape-RNase HⅡ exhibited the highest activity in the presence of 5 mM Mn2+, 1 mM Co2+, or 10mM Mg2+, respectively;however,its cleavage efficiencies at different cleavage sites for Mn2+ and Mg2+ 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 DNAr4-DNA/DNA and 13-bp DNA-r1-DNA/DNA chimeric substrates were cleaved at DNA-RNA junction.ApeRNase 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 ApeRNase HⅡ and the results should contribute to the analysis of RNase H of other archaeal species.