Identification by Phage Display Selection of a Short Peptide Able To Inhibit Only the Strand Transfer Reaction Catalyzed by Human Immunodeficiency Virus Type 1 Integrase

Desjobert, Cécile; Soultrait, Vaea Richard de; Faure, A.; Parissi, Vincent; Litvak, Simón; Tarragó-Litvak, Laura; Fournier, Michel

doi:10.1021/bi049385e

Cited by 35 publications

(34 citation statements)

References 40 publications

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“…Based on the observation that this peptide also inhibited the IN from HIV-2, FIV, and MLV, it was suggested that a conserved region around the catalytic domain of IN is being targeted. An IN inhibitory peptide was also selected using a phage display library (37). IN-derived peptides that interfered with its oligomerization also blocked its enzymatic activity (38).…”

Section: The Rev-derived Short Peptides (Rev13-23 and Rev53-67) Inhibmentioning

confidence: 99%

Interaction between HIV-1 Rev and Integrase Proteins

Rosenbluh

Hayouka

Loya

et al. 2007

Journal of Biological Chemistry

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Following penetration of human immunodeficiency virus 1 (HIV-1) 3 into the host cell, reverse transcription of the viral RNA produces cDNA that is then integrated into the host chromosome (1). The integration of the viral cDNA is an essential early step in the HIV-1 life cycle. This reaction is catalyzed by the viral integrase (IN), a 32-kDa protein that is an integral part of the viral pre-integration complex (PIC) (2). The IN protein is encoded by the viral pol gene and is translated as part of a large Gag-Pol polyprotein that is processed by the viral protease (3, 4).For the integration process to occur, the viral IN must recognize specific sequences in the viral cDNA, at the termini of the long terminal repeat (LTR) elements. Retroviral integration proceeds in two steps: in the first, termed 3Ј-end processing, a dinucleotide is removed from the 3Ј-end. This reaction occurs in the cytoplasm, within the PIC (5, 6). In the next step, after entering the nucleus, the processed viral double-stranded DNA is joined to the host target DNA by an IN-mediated strandtransfer reaction.Due to its central role in HIV replication, the IN protein is an attractive target for antiviral therapy (4). Identifying specific IN inhibitors may provide a novel approach for multi-therapy strategies. Moreover, probably no cellular counterpart of IN exists in human cells and therefore, IN inhibitors should not interfere with normal cellular processes. However, only a few IN inhibitors have been described to date (4). This is in contrast to the number of inhibitors obtained against reverse transcriptase (RT) and the protease, which are the other HIV-1 enzymes that are currently used as targets for the development of anti-HIV-1 drugs (4). A promising approach for the discovery of IN inhibitors is the use of peptides derived from the IN-binding sequences of IN-interacting proteins.Specific domains within viral proteins are responsible for their interaction with host-cell receptors and with other viral and cellular proteins enabling the completion of the viral propagation cycle within the host cell (1, 6). Peptides derived from these binding domains may interfere with virus-host and virusvirus protein interactions and as such are excellent candidates as therapeutic agents. Using this approach, short peptides that inhibit IN enzymatic activity were obtained following analysis of the interaction between two of the HIV-1 proteins, RT and IN. Screening a complete library of RT-derived peptides demonstrated that two domains of about 20 amino acids mediate this interaction. Peptides bearing these amino acid sequences blocked IN enzymatic activities in vitro (7).In the present work, we observed a not yet described interaction between the HIV-1 Rev and IN proteins. The HIV-1 Rev is a karyophilic protein, which is required at the late phase of the viral life cycle for promoting nuclear export of partially spliced or un-spliced viral RNA (8, 9). Based on this finding, we identi-* This work was supported in part by grants from the Israel Science Foundat...

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Section: The Rev-derived Short Peptides (Rev13-23 and Rev53-67) Inhibmentioning

confidence: 99%

Interaction between HIV-1 Rev and Integrase Proteins

Rosenbluh

Hayouka

Loya

et al. 2007

Journal of Biological Chemistry

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“…proteins (41), and peptides. The peptides found to be active against HIV-1 IN are those derived from sequences within IN (42)(43)(44)(45), from random sequences identified in synthetic libraries (45)(46)(47), from combinatorial libraries of chemically synthesized peptides (45,48), or from cellular IN-interacting proteins (45,49,50). In contrast to many anti-RT drugs used in the treatment of AIDS patients, there is only one group of nonpeptide anti-IN drugs that has entered clinical trials (39,51), and none is used routinely so far in patients.…”

Section: Hiv-1 Rt and Rt-derived Peptides 4286 And 4321 Physically Inmentioning

confidence: 99%

Peptides Derived from the Reverse Transcriptase of Human Immunodeficiency Virus Type 1 as Novel Inhibitors of the Viral Integrase

Gleenberg

Avidan

Goldgur

et al. 2005

Journal of Biological Chemistry

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Two viral encoded enzymes play central roles in the early stages of the replication of retroviruses and retrotransposons. The first one, reverse transcriptase (RT), 1 converts the singlestranded viral RNA into double-stranded DNA in a relatively complex process, reverse transcription. This step is catalyzed by the two catalytic activities of RT, the DNA polymerase (capable of copying both RNA and DNA into DNA) and the RNase H activity, which concomitantly hydrolyzes the RNA strand in the DNA-RNA heteroduplex formed (1). Subsequently, the RT-produced double-stranded DNA is transported into the nucleus, as part of the nucleoprotein complex (designated the preintegration complex), where it integrates into the genomic target DNA by the second viral enzyme, the IN. IN identifies the ends of the linear viral DNA, trims them (by removing two or three extra nucleotides located 3Ј to the highly conserved CA 3Ј termini), and then accompanies the DNA into the nucleus to catalyze integration into the target cellular DNA (1-3). There are several examples for potential linkages between RT and IN. First, the DNA product of RT is the substrate for IN, the next enzyme in the line of the viral replication cycle. Second, both proteins are proteolytic products of the same polyprotein precursor encoded by a single retroviral gene, the pol (1). In some cases, as in avian sarcoma leukosis virus, the IN sequence appears in two forms, one as part of the large ␤-subunit of the RT and the other as a free IN protein designated pp32 (1, 4). Moreover, PICs, which are capable of performing in vitro integration, contain the viral DNA, IN, RT, and other proteins (5-8). Third, the INs and RTs of HIV-1 and MLV were shown to exhibit physical interactions (9 -11). These direct contacts between the IN and RT of HIV-1 were recently confirmed by us by using surface plasmon resonance technology. 2 Finally, we and others (12, 13) have shown recently that RT can inhibit in vitro the enzymatic activities of IN, suggesting functional roles for these interactions.After the completion of reverse transcription in the cytoplasm of the infected cell, there is a significant delay in the process of integration depending on the rate by which the PICs are transported into the nucleus (1). Because all possible catalytic components for integration are likely to be present in the PICs, the viral DNA can serve as donor DNA as well as the target DNA for integration. Such a potential auto-integration process is suicidal for the virus, as it destructs the viral genome. For that reason, it is imperative to understand the mechanisms that regulate the integration of the retroviral genome and learn how to control it. Several cellular proteins are known to be involved in the integration process in HIV-infected cells (14 -17). Most interestingly, one of these factors serves also as a barrier to auto-integration in MLV-infected cells (18).

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“…Generally, the IC 50 for peptides selected from phage display are in the nM to μM range. 27,28 Compared with these peptides, the low IC 50 for peptide T3-15 is likely due to the selection method and longer elution conditions established here, and may also indicate that it has a higher binding affinity for the TAG-72 over the peptides selected by us for tumor thigh/normal thigh and tumor thigh/blood from 5-90 min. SPECT/CT imaging was performed in another set of mice receiving 20 μg peptide radiolabeled with 200 μCi with acquisitions started at 20 and 60 min (Fig.…”

Section: Resultsmentioning

confidence: 94%