Crystal Structures of the Reverse Transcriptase-associated Ribonuclease H Domain of Xenotropic Murine Leukemia-virus Related Virus

Overview

Journal J Struct Biol

Specialties Cell Biology
Molecular Biology

Date 2012 Feb 28

PMID 22366278

Citations 10

Authors

Dongwen Zhou

Suhman Chung

Maria Miller

Stuart F J Le Grice

Alexander Wlodawer

Affiliations

Soon will be listed here.

Abstract

The ribonuclease H (RNase H) domain of retroviral reverse transcriptase (RT) plays a critical role in the life cycle by degrading the RNA strands of DNA/RNA hybrids. In addition, RNase H activity is required to precisely remove the RNA primers from nascent (-) and (+) strand DNA. We report here three crystal structures of the RNase H domain of xenotropic murine leukemia virus-related virus (XMRV) RT, namely (i) the previously identified construct from which helix C was deleted, (ii) the intact domain, and (iii) the intact domain complexed with an active site α-hydroxytropolone inhibitor. Enzymatic assays showed that the intact RNase H domain retained catalytic activity, whereas the variant lacking helix C was only marginally active, corroborating the importance of this helix for enzymatic activity. Modeling of the enzyme-substrate complex elucidated the essential role of helix C in binding a DNA/RNA hybrid and its likely mode of recognition. The crystal structure of the RNase H domain complexed with β-thujaplicinol clearly showed that coordination by two divalent cations mediates recognition of the inhibitor.

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