Modeling the Late Steps in HIV-1 Retroviral Integrase-catalyzed DNA Integration

Brin, Elena; Yi, Jizu; Skalka, Anna Marie; Leis, Jonathan

doi:10.1074/jbc.m006929200

Cited by 60 publications

(46 citation statements)

References 32 publications

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“…The process is completed by the third main step, gap repair, yielding a duplication of host DNA flanking the provirus (e.g., the five dark shaded bases shown here for HIV). Like the first two steps [6,11,13,14,195], gap repair can also be modeled in vitro [196,197]. It appears to involve host cell DNA repair systems [10].…”

Section: Targeting Lentiviral Vectorsmentioning

confidence: 99%

Integrase, LEDGF/p75 and HIV replication

Poeschla

2008

Cell. Mol. Life Sci.

115

104

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HIV integrates a DNA copy of its genome into a host cell chromosome in each replication cycle. The essential DNA cleaving and joining chemistry of integration is known, but there is less understanding of the process as it occurs in a cell, where two complex and dynamic macromolecular entities are joined: the viral pre-integration complex and chromatin. Among implicated cellular factors, much recent attention has coalesced around LEDGF/p75, a nuclear protein that may act as a chromatin docking factor or receptor for lentiviral pre-integration complexes. LEDGF/p75 tethers HIV integrase to chromatin, protects it from degradation, and strongly influences the genome-wide pattern of HIV integration. Depleting the protein from cells and/or over-expressing its integrase-binding domain blocks viral replication. Current goals are to establish the underlying mechanisms and to determine whether this knowledge can be exploited for antiviral therapy or for targeting lentiviral vector integration in human gene therapy.

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Section: Targeting Lentiviral Vectorsmentioning

confidence: 99%

Integrase, LEDGF/p75 and HIV replication

Poeschla

2008

Cell. Mol. Life Sci.

115

104

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“…IN inserts the newly processed 3Ј ends of the viral DNA into the host genome in a coupled cleavage-ligation reaction called 3Ј-end joining or strand transfer. Host enzymes are believed to carry out the repair of gaps between viral and host DNAs after strand transfer (6,14,60), resulting in the formation of a provirus.…”

mentioning

confidence: 99%

Requirement for Integrase during Reverse Transcription of Human Immunodeficiency Virus Type 1 and the Effect of Cysteine Mutations of Integrase on Its Interactions with Reverse Transcriptase

Zhu¹,

Dobard²,

Chow³

2004

J Virol

143

161

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Retroviral integrase catalyzes the essential step of integrating a double-stranded DNA copy of the viral genome into a host cell chromosome. Mutational studies have revealed that integrase is involved in additional steps of viral replication, but the mechanism for the pleiotropic effect is not well characterized. Since Cys residues generally play crucial roles in protein structure and function, we introduced Cys-to-Ser substitutions at positions 56, 65, and 130 of human immunodeficiency virus type 1 (HIV-1) integrase to determine their effects on integration activity and viral replication. None of the substitutions significantly affected the enzymatic activities in vitro. When introduced into the NL4-3 molecular clone of HIV-1, mutant viruses encoding Cys mutations at positions 56 and 65 of integrase replicated similarly to the wild-type virus in CD4؉ -T-cell lines, whereas the C130S-containing virus was noninfectious. The entry and postintegration steps of the viral life cycle for all mutant viruses were normal, and all had particle-associated reverse transcriptase (RT) activity. However, early reverse-transcribed DNA products were absent in the lysate of cells infected with the C130S mutant virus, indicating that the mutation abolished the ability of the virus to initiate endogenous reverse transcription. Coimmunoprecipitation using purified integrase and RT showed that the C-terminal domain of wild-type HIV-1 integrase interacted with RT. The interaction between integrase and RT was not affected in the presence of a reducing or alkylating agent, suggesting that the interaction did not involve a disulfide linkage. The C130S substitution within the core region may disrupt the protein recognition interface of the C-terminal domain and abolish its ability to interact with RT. Our results indicate that integrase plays an important role during the reverse-transcription step of the viral life cycle, possibly through physical interactions with RT.

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“…This is a concerted DNA cleavage-ligation reaction whereby the host DNA is cleaved and both 3Ј ends of the viral DNA are covalently linked to the 5Ј ends of the host DNA. Repair of the 5-base single-stranded DNA gaps between the 5Ј ends of the viral DNA and the 3Ј ends of the host DNA is likely carried out by host cell enzymes (4,49). A possible role of integrase in DNA splicing (13) has also been proposed.…”

mentioning

confidence: 99%

Impact of the Central Polypurine Tract on the Kinetics of Human Immunodeficiency Virus Type 1 Vector Transduction

Maele

Rijck

Clercq

et al. 2003

J Virol

121

107

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In contrast to oncoretroviruses, human immunodeficiency virus type 1 (HIV-1) and other lentiviruses have the capacity to replicate in nondividing cells such as macrophages. Lentiviral integration requires transport of the preintegration complex (PIC) through the nuclear pores by an active, energy-dependent process (6). At least three components of the HIV-1 PIC seem to contribute to nuclear transport: Vpr (23,27,28,42,45), matrix protein (7, 24, 48), and integrase (3, 25, 38). These different proteins interact with each other and with the cellular import machinery (14). However, the respective contribution of each protein to the karyophilic properties of HIV-1 PICs has not been completely elucidated (17,18,22,31). Nuclear import is not determined by viral proteins only. Recently, it was shown that the central DNA flap of HIV-1 acts as a cis determinant of HIV-1 DNA nuclear import (21, 51). During the second-strand DNA synthesis of reverse transcription, initiation of DNA synthesis at the central polypurine tract (cPPT) results in a strand displacement event. The cPPT, an exact copy of the PPT at the 3Ј end, serves as the second initiation site of second-strand DNA synthesis, while the central termination sequence (CTS) marks the termination of strand displacement synthesis. The final product of HIV-1 reverse transcription is a linear DNA molecule with a central 99-nucleotide-long plusstrand overlap, the central DNA flap (10, 11). The central DNA flap is believed to act as a cis determinant for HIV-1

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Modeling the Late Steps in HIV-1 Retroviral Integrase-catalyzed DNA Integration

Cited by 60 publications

References 32 publications

Integrase, LEDGF/p75 and HIV replication

Integrase, LEDGF/p75 and HIV replication

Requirement for Integrase during Reverse Transcription of Human Immunodeficiency Virus Type 1 and the Effect of Cysteine Mutations of Integrase on Its Interactions with Reverse Transcriptase

Impact of the Central Polypurine Tract on the Kinetics of Human Immunodeficiency Virus Type 1 Vector Transduction

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