Proteolytic release and crystallization of the RNase H domain of human immunodeficiency virus type 1 reverse transcriptase.
The RNase H domain of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase was released from recombinant DHFR-RNase H fusion protein by the action of HIV-1 protease and crystallized as large trigonal prisms that diffract x-rays to at least 2.4-A resolution. The protease cleavage occurred 18 residues away from the Phe440-Tyr441 site reported to be processed during maturation of the reverse transcriptase heterodimer. Mutagenesis of the protease-sensitive region (residues 430-440), which is part of the crystallized domain, indicates that any alteration of the wild-type sequence results in increased proteolysis of the p66 subunit. A model of asymmetric processing in HIV-1 reserve transcriptase which involves partial unfolding of the RNase H domain is proposed based on these results and the recently reported three-dimensional structure of this domain.
Hostomska
Z
Z
Agouron Pharmaceuticals, Inc., La Jolla, California 92037.
Matthews
D A
DA
Davies
J F
JF
Nodes
B R
BR
Hostomsky
Z
Z
D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000737:chemistry*; D015497:HIV-1 / Q000201:enzymology*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry*; D016914:Ribonuclease H; D014961:X-Ray Diffraction
D012194:RNA-Directed DNA Polymerase; D004722:Endoribonucleases; D016914:Ribonuclease H