Analysis of Nucleoside Triphosphate Hydrolysis by Middle East Respiratory Syndrome Coronavirus Helicase

초록

The purpose of this study is to measure the nucleoside triphosphate (NTP) hydrolysis by Middle East Respiratory Syndrome coronavirus (MERS-CoV) helicase in the presence of different types of nucleic acids (NAs) for the determination of the optimal condition of hydrolysis activity. The steady-state NTP hydrolysis analysis demonstrated that the MERS-CoV helicase only (and considerably) hydrolyzed the ATP in the presence of poly(U). On the contrary, only UTP was considerably hydrolyzed in the presence of poly(C). When we used the DNAs as a substrate, the single-stranded M13 DNA considerably stimulated the ATP and UTP hydrolysis. However, the ATP hydrolysis was not detected in the presence of ds DNA. The pH-dependent ATP hydrolysis analysis demonstrated that the helicase optimally hydrolyzed ATP at a pH value of 7.0. The ATP hydrolysis was also performed in the presence of four different kinds of divalent cations, which demonstrated that only Mg2+ ions stimulated the ATP hydrolysis activity.

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