Biochemical Characterization of Novel Retroviral Integrase Proteins

Ballandras-Colas, Allison; Naraharisetty, Hema L.; Li, Xiang; Serrao, Erik; Engelman, Alan

doi:10.1371/journal.pone.0076638

Cited by 17 publications

(30 citation statements)

References 36 publications

(53 reference statements)

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“…94−101 Conversely, some recombinant IN proteins, in particular PFV IN, are far more proficient at full-site strand transfer. 102−104 The underlying reasons for these differences between divergent IN proteins are not fully understood. The propensity of HIV-1 IN to self-associate into higher-order multimers in solution 105 could be a factor limiting its concerted integration activity in vitro.…”

Section: Enzymatic Steps In Retroviral Dna Integrationmentioning

confidence: 99%

Retroviral DNA Integration

2016

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The integration of a DNA copy of the viral RNA genome into host chromatin is the defining step of retroviral replication. This enzymatic process is catalyzed by the virus-encoded integrase protein, which is conserved among retroviruses and LTR-retrotransposons. Retroviral integration proceeds via two integrase activities: 3′-processing of the viral DNA ends, followed by the strand transfer of the processed ends into host cell chromosomal DNA. Herein we review the molecular mechanism of retroviral DNA integration, with an emphasis on reaction chemistries and architectures of the nucleoprotein complexes involved. We additionally discuss the latest advances on anti-integrase drug development for the treatment of AIDS and the utility of integrating retroviral vectors in gene therapy applications.

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Section: Enzymatic Steps In Retroviral Dna Integrationmentioning

confidence: 99%

Retroviral DNA Integration

2016

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“…Such differences contribute to the features that distinguish their behaviors in solution. IN proteins from several retroviruses differ in both solubility and multimerization properties (91, 92). For example, in the absence of DNA and at similar protein concentration (~60 µM), ASLV IN is a dimer and HIV-1 IN is a tetramer, whereas PFV IN is a monomer even at twice this concentration (93).…”

Section: Integrase Structurementioning

confidence: 99%

Retroviral Integrase: Then and Now

Andrake

Skalka

2015

Annu. Rev. Virol.

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The retroviral integrases are virally encoded, specialized recombinases that catalyze the insertion of viral DNA into the host cell’s DNA, a process that is essential for virus propagation. We have learned a great deal since the existence of an integrated form of retroviral DNA (the provirus) was first proposed by Howard Temin in 1964. Initial studies focused on the genetics and biochemistry of avian and murine virus DNA integration, but the pace of discovery increased substantially with advances in technology, and an influx of investigators focused on the human immunodeficiency virus (HIV). We begin with a brief account of the scientific landscape in which some of the earliest discoveries were made, and summarize research that led to our current understanding of the biochemistry of integration. A more detailed account of recent analyses of integrase structure follows, as they have provided valuable insights into enzyme function and raised important new questions.

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“…Epsilonretrovirus integrase was reportedly insoluble following its expression in Escherichia coli under a variety of conditions (55). HIV-1 integrase purified following its expression in bacteria can attain concentrations of 1 mg/ml or greater, however this strictly depends on nonphysiological concentrations of salt (e.g., 1 M NaCl).…”

Section: Challenges Of Working With Recombinant Integrase Proteinsmentioning

confidence: 99%

“…Initially recognized as a DX [39][40][41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56][57][58] DX 35 E motif conserved among retroviral and retrotransposon integrases and bacterial IS3 insertion sequences (74,78,81), the advent of genomic sequencing has since expanded the DD(E/D) superfamily of polynucleotidyl transferases to include the transposase proteins of numerous prokaryotic and eukaryotic transposable elements (47,(82)(83)(84). To date, the roles of the active site residues during integrative recombination are most thoroughly understood from X-ray crystallographic analyses of active PFV integrase-DNA complexes (85).…”

Section: Integrase Protein Structures Domain Organization Of Integrasmentioning

confidence: 99%

Retroviral Integrase Structure and DNA Recombination Mechanism

Engelman

Cherepanov

2014

Microbiol Spectr

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SUMMARYDue to the importance of human immunodeficiency virus type 1 (HIV-1) integrase as a drug target, the biochemistry and structural aspects of retroviral DNA integration have been the focus of intensive research during the past three decades. The retroviral integrase enzyme acts on the linear double-stranded viral DNA product of reverse transcription. Integrase cleaves specific phosphodiester bonds near the viral DNA ends during the 3′ processing reaction. The enzyme then uses the resulting viral DNA 3′-OH groups during strand transfer to cut chromosomal target DNA, which simultaneously joins both viral DNA ends to target DNA 5′-phosphates. Both reactions proceed via direct transesterification of scissile phosphodiester bonds by attacking nucleophiles: a water molecule for 3′ processing, and the viral DNA 3′-OH for strand transfer. X-ray crystal structures of prototype foamy virus integrase-DNA complexes revealed the architectures of the key nucleoprotein complexes that form sequentially during the integration process and explained the roles of active site metal ions in catalysis. X-ray crystallography furthermore elucidated the mechanism of action of HIV-1 integrase strand transfer inhibitors, which are currently used to treat AIDS patients, and provided valuable insights into the mechanisms of viral drug resistance.

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Biochemical Characterization of Novel Retroviral Integrase Proteins

Cited by 17 publications

References 36 publications

Retroviral DNA Integration

Retroviral DNA Integration

Retroviral Integrase: Then and Now

Retroviral Integrase Structure and DNA Recombination Mechanism

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