Crystallographic analyses of an active HIV-1 ribonuclease H domain show structural features that distinguish it from the inactive form

Chattopadhyay, Debasish; Finzel, B.C.; Munson, Sibyl H.; Evans, David B.; Sharma, Satish Kumar; Strakalaitus, N.A; Brunner, David P.; Eckenrode, F.M; Dauter, Zbigniew; Betzel, Christian; Einspahr, Howard

doi:10.1107/s0907444993002409

Cited by 19 publications

(26 citation statements)

References 13 publications

(24 reference statements)

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“…Several new sequences have been added to the database of substrates for the HIV-1 protease and these serve to reinforce past generalizations with regard to preferences of the enzyme for amino acids in particular P-sites (Poorman et al, 1991;Chou, 1993;Chou et al, 1993). Our results, interpreted with respect to recently published X-ray crystallographic structures for RNase H (Davies et al, 1991;Hostomska et al, 1991 ;Chattopadhyay et al, 1993) and p66/p51 (Kohlstaedt et al, 1992), are consistent with the view that the RT homodimer adopts a conformation similar to that of the heterodimeric p66/p5 1, but in which the COOH-terminal RNase H domain of one protomer is disordered.…”

mentioning

confidence: 59%

“…Based upon the structural analysis, Davies and coworkers (1991) noted that the Phe,,,, cleavage site that transforms the p66/p66 homodimer to p66/p51 is in a @sheet region and is inaccessible to protease binding. This is illustrated in Figure 5 and Kinemage 1, based upon a recently determined structure of our RNase H construct (Chattopadhyay et al, 1993). Davies et al (1991) reasoned that the p66/p66 homodimer would be asymmetric; one RNase H domain would be properly folded and refractive to protease excision and the other would be at least partially unfolded to allow cleavage at the Phe-…”

Section: Discussionmentioning

confidence: 99%

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Human immunodeficiency virus type‐1 reverse transcriptase and ribonuclease h as substrates of the viral protease

Tomasselli¹,

Sarcich²,

Barrett³

et al. 1993

Protein Science

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A study has been made of the susceptibility of recombinant constructs of reverse transcriptase (RT) and ribonuclease H (RNase H) from human immunodeficiency virus type 1 (HIV-1) to digestion by the HIV-1 protease. At neutral pH, the protease attacks a single peptide bond, Phe440-Tyr441, in one of the protomers of the folded, active RT/RNase H (p66/p66) homodimer to give a stable, active heterodimer (~6 6 1~5 1 ) that is resistant to further hydrolysis (Chattopadhyay, D., et al., 1992, J. Biol. Chem. 267, 14227-14232). The COOH-terminal p15 fragment released in the process, however, is rapidly degraded by the protease by cleavage at Tyr,,,-Leu,,, and Leu,,,. In marked contrast to this p15 segment, both p66/p51 and a folded RNase H construct are stable to breakdown by the protease at neutral pH. It is only at pH values around 4 that these latter proteins appear to unfold and, under these conditions, the heterodimer undergoes extensive proteolysis. RNase H is also hydrolyzed at low pH, but cleavage takes place primarily at Gly,,,-Ala,,, and at Phe440-Tyr441, and only much more slowly at residues 483,494, and 532. This observation can be reconciled by inspection of crystallographic models of RNase H, which show that residues 483,494, and 532 are relatively inaccessible in comparison to Gly,,, and Phe,,. Our results fit a model in which the p66/p66 homodimer exists in a conformation that mirrors that of the heterodimer, but with a plS segment on one of the protomers that is structurally disordered to the extent that all of its potential HIV protease cleavage sites are accessible for hydrolysis.

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mentioning

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Human immunodeficiency virus type‐1 reverse transcriptase and ribonuclease h as substrates of the viral protease

Tomasselli¹,

Sarcich²,

Barrett³

et al. 1993

Protein Science

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“…Crystals of one of these active domains have been obtained (an N-terminally immobilized metal-affinity chromatography-tagged HIV RNase H), and preliminary data suggest that the short His-containing loop is ordered in this active mutant (34). The metal-affinity tagged domains lack the basic helix/loop region seen in E. coli.…”

Section: Discussionmentioning

confidence: 99%

Substitution of a highly basic helix/loop sequence into the RNase H domain of human immunodeficiency virus reverse transcriptase restores its Mn(2+)-dependent RNase H activity.

Keck

Marqusee

1995

Proc. Natl. Acad. Sci. U.S.A.

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“…Two crystal structures of the isolated RNase H domain of HIV-1 RT have been published (PDB code 1HRH, (Davies et al, 1991); and PDB code 1RDH (Chattopadhyay et al, 1993)), as well as a solution (NMR) structure (Pari et al, 2003). The most structurally detailed of the isolated HIV-1 RT RNase H domains is that of PDB file 1HRH.…”

Section: Momlv Rt Rnase H and Hiv-1 Rnase Hmentioning

confidence: 99%

Murine leukemia virus reverse transcriptase: Structural comparison with HIV-1 reverse transcriptase

Coté¹,

Roth²

2008

Virus Research

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Recent X-ray crystal structure determinations of Moloney murine leukemia virus reverse transcriptase (MoMLV RT) have allowed for more accurate structure/function comparisons to HIV-1 RT than were formerly possible. Previous biochemical studies of MoMLV RT in conjunction with knowledge of sequence homologies to HIV-1 RT and overall fold similarities to RTs in general, provided a foundation upon which to build. In addition, numerous crystal structures of the MoMLV RT fingers/palm subdomain had also shed light on one of the critical functions of the enzyme, specifically polymerization. Now in the advent of new structural information, more intricate examination of MoMLV RT in its entirety can be realized, and thus the comparisons with HIV-1 RT may be more critically elucidated. Here, we will review the similarities and differences between MoMLV RT and HIV-1 RT via structural analysis, and propose working models for the MoMLV RT based upon that information.

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Crystallographic analyses of an active HIV-1 ribonuclease H domain show structural features that distinguish it from the inactive form

Cited by 19 publications

References 13 publications

Human immunodeficiency virus type‐1 reverse transcriptase and ribonuclease h as substrates of the viral protease

Human immunodeficiency virus type‐1 reverse transcriptase and ribonuclease h as substrates of the viral protease

Substitution of a highly basic helix/loop sequence into the RNase H domain of human immunodeficiency virus reverse transcriptase restores its Mn(2+)-dependent RNase H activity.

Murine leukemia virus reverse transcriptase: Structural comparison with HIV-1 reverse transcriptase

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