Insight into the Mechanism of a Peptide Inhibitor of HIV Reverse Transcriptase Dimerization

Depollier, Julien; Hourdou, Marie-Laure; Aldrian, Gudrun; Rothwell, Paul J.; Restle, Tobias; Divita, Gilles

doi:10.1021/bi0484264

Cited by 28 publications

(44 citation statements)

References 38 publications

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“…They are found to play critical roles in viral DNA replication, in fidelity, and in drug resistance (Agopian et al, 2007;Dash et al, 2006;Depollier et al, 2005;Kim et al, 1998Kim et al, , 1999.…”

Section: Discussionmentioning

confidence: 99%

Mechanistic insights into the roles of three linked single-stranded template binding residues of MMLV reverse transcriptase in misincorporation and mispair extension fidelity of DNA synthesis

Xie

Zhang

Li³

et al. 2011

Gene

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Section: Discussionmentioning

confidence: 99%

Mechanistic insights into the roles of three linked single-stranded template binding residues of MMLV reverse transcriptase in misincorporation and mispair extension fidelity of DNA synthesis

Xie

Zhang

Li³

et al. 2011

Gene

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“…wiley.com. ] dimeric conformation of the enzyme [Depollier et al, 2005]. Thus, targeting dimer stability or disruption of dimerization of HIV-1 RT provides an attractive approach for intervention in HIV-1 replication.…”

Section: Discussionmentioning

confidence: 99%

A single deletion at position 134, 135, or 136 in the beta 7–beta 8 loop of the p51 subunit of HIV‐1 RT disrupts the formation of heterodimeric enzyme

Upadhyay

Pandey

Mishra

et al. 2009

J of Cellular Biochemistry

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The human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is a heterodimeric enzyme composed of p66 and p51 subunits. Earlier, we showed that the beta7-beta8 loop of p51 is crucial for polymerase activity of HIV-1 RT as either deletion or Ala substitution of amino acids in the beta7-beta8 loop spanning residues 136-139 in the p51 subunit impaired dimerization and, in turn, polymerase function of the enzyme (Pandey et al. 2001 Biochemistry 40: 9505-9512). In the present study, we generated subunit-specific single-deletion mutants at positions 134, 135, 136, or 137 and examined their effects on the heterodimerization, binary complex formation, and polymerase functions of the enzyme. We found that among these four residues, Ser134, Ile135, and Asn136 in the beta7-beta8 loop of the p51 subunit are crucial residues for dimerization and polymerase function of the enzyme, but have no impact when specifically deleted from the p66 subunit. These results demonstrate the beta7-beta8 loop of the p51 subunit in the formation of stable, functional heterodimeric enzyme which could be an attractive target for anti-HIV-1 drug development.

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“…In protein-protein interactions the binding energy is not evenly distributed across the dimer interface but involves specific residues "hot spots" that stabilize protein complexes. We have shown that the use of small peptides targeting hot spot residues required for RT dimerization constitutes a new strategy to inhibit HIV-1 RT (16, 17) and have described a decapeptide "Pep-7"-mimicking p66/p51 interface that prevents RT dimerization by destabilizing RT subunit interactions and that blocks viral replication (23,32).…”

Section: Human Immunodeficiency Virus Type I (Hiv-1)mentioning

confidence: 99%

“…Expression and Purification of HIV-1 RT Proteins-Histagged RTs were expressed and purified as previously described (23,34). Briefly, M15 bacteria (Qiagen) were separately transformed with all the constructs of p51 and p66 subunits.…”

Section: Materials-poly(ra)-oligo(dt) and [mentioning

confidence: 99%

A New Generation of Peptide-based Inhibitors Targeting HIV-1 Reverse Transcriptase Conformational Flexibility

Agopian

Gros

Aldrian

et al. 2009

Journal of Biological Chemistry

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The biologically active form of human immunodeficiency virus (HIV) type 1 reverse transcriptase (RT) is a heterodimer. The formation of RT is a two-step mechanism, including a rapid protein-protein interaction "the dimerization step," followed by conformational changes "the maturation step," yielding the biologically active form of the enzyme. We have previously proposed that the heterodimeric organization of RT constitutes an interesting target for the design of new inhibitors. Here, we propose a new class of RT inhibitors that targets protein-protein interactions and conformational changes involved in the maturation of heterodimeric reverse transcriptase. Based on a screen of peptides derived from the thumb domain of this enzyme, we have identified a short peptide P AW that inhibits the maturation step and blocks viral replication at subnanomolar concentrations. P AW only binds dimeric RT and stabilizes it in an inactive/non-processive conformation. From a mechanistic point of view, P AW prevents proper binding of primer/template by affecting the structural dynamics of the thumb/fingers of p66 subunit. Taken together, these results demonstrate that HIV-1 RT maturation constitutes an attractive target for AIDS chemotherapeutics. Human immunodeficiency virus type I (HIV-1)4 is the primary cause of AIDS, a slow progressive and degenerative disease of the human immune system. Despite recent therapeutic developments and the introduction of highly active antiretroviral therapy, the rapid emergence of drug-resistant viruses against all approved drugs together with inaccessible latent virus reservoirs and side effects of currently used compounds have limited the efficacy of existing anti-HIV-1 therapeutics (1).Therefore, there is still an urgent need for new and safer drugs, active against resistant viral strains or directed toward novel targets in the replicative cycle, which will be useful for multiple drug combination.HIV-1 reverse transcriptase (RT) plays an essential multifunctional role in the replication of the virus, by catalyzing the synthesis of double-stranded DNA from the single strand retroviral RNA genome (2, 3). The majority of the chemotherapeutic agents used in AIDS treatments target the polymerase activity of HIV-1 RT, such as nucleoside reverse transcriptase inhibitors (NRTI) or non-nucleoside inhibitors (NNRTI) (4). The biologically active form of RT is an asymmetric heterodimer that consists of two subunits, p66 and p51, derived from p66 by proteolytic cleavage of the C-terminal RNase H domain (2, 3, 5).The polymerase domain of both p66 and p51 subunit can be subdivided into four common subdomains: fingers, palm, thumb, and connection (6 -10). Determination of the threedimensional structures of RTs has revealed that, although the folding of individual subdomains is similar in p66 and p51, their spatial arrangement differs markedly (11). The p66 subunit contains both polymerase and RNase H active sites. The p66-polymerase domain folds into an "open," extended structure, forming a large active site ...

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Insight into the Mechanism of a Peptide Inhibitor of HIV Reverse Transcriptase Dimerization

Cited by 28 publications

References 38 publications

Mechanistic insights into the roles of three linked single-stranded template binding residues of MMLV reverse transcriptase in misincorporation and mispair extension fidelity of DNA synthesis

Mechanistic insights into the roles of three linked single-stranded template binding residues of MMLV reverse transcriptase in misincorporation and mispair extension fidelity of DNA synthesis

A single deletion at position 134, 135, or 136 in the beta 7–beta 8 loop of the p51 subunit of HIV‐1 RT disrupts the formation of heterodimeric enzyme

A New Generation of Peptide-based Inhibitors Targeting HIV-1 Reverse Transcriptase Conformational Flexibility

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