Rous Sarcoma Virus Synaptic Complex Capable of Concerted Integration Is Kinetically Trapped by Human Immunodeficiency Virus Integrase Strand Transfer Inhibitors

Pandey, Krishan K.; Bera, Sibes; Korolev, Sergey; Campbell, Mary Ann; Yin, Zhiqi; Aihara, Hideki; Grandgenett, Duane P.

doi:10.1074/jbc.m114.573311

Cited by 8 publications

(64 citation statements)

References 43 publications

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“…The subunit structure of IN is varied and highly dependent on the purification process and different salt requirements to maintain solubility for each expressed recombinant IN. PFV IN is soluble (10-15 mg/mL) in 0.2 mol/L NaCl [48,49] while RSV IN is highly soluble (30 mg/mL) in 0.2 mol/L (NH4)2SO4 [50] . PFV IN is monomeric [49] while RSV IN is a dimer in solution [50][51][52] .…”

Section: Solution Properties Of Inmentioning

confidence: 99%

“…PFV IN is soluble (10-15 mg/mL) in 0.2 mol/L NaCl [48,49] while RSV IN is highly soluble (30 mg/mL) in 0.2 mol/L (NH4)2SO4 [50] . PFV IN is monomeric [49] while RSV IN is a dimer in solution [50][51][52] . Recombinant HIV IN has been purified as a monomer, dimer and tetramer [47,53,54] .…”

Section: Solution Properties Of Inmentioning

confidence: 99%

“…The HIV IN monomer purified in the presence of EDTA is converted into a tetramer in the presence of Zn 2+ without the formation of a dimer intermediate [53,54] . Studies have demonstrated that a tetramer of IN is associated with HIV, PFV and RSV SC capable of concerted integration [50,[55][56][57] . The oligomeric state of retrovirus IN play key roles in virus maturation upon release of the virus particle, enzymatic activities for 3' OH processing and concerted integration of viral DNA and possibly for reverse transcription in the cytoplasmic PIC.…”

Section: Solution Properties Of Inmentioning

confidence: 99%

“…There are strong structural similarities in the CCD between HIV, PFV and RSV IN (Figure 2) [47,51,52,57,67,76,77] suggesting that under appropriate conditions the highly soluble RSV IN could also be trapped in the presence of the appropriate oligonucleotide (ODN) substrate and STIs. Assembly of a trapped RSV SC at high IN concentrations (1.5 mg/mL; 45 µmol/L) in solution required the presence of 3' OH recessed viral ODN ranging in sizes (16/18R to 18/20R) and DTG, RAL or MK-2048 [50] (Figure 9). DTG and MK-2048 (an investigational inhibitor) possess a significantly longer dissociative half-life than RAL within an HIV IN-DNA complex allowing these STIs to capture and stabilize the IN-DNA complex [78,79] .…”

Section: Hiv and Rsv Synaptic Complexes Are Kinetically Stabilized Bymentioning

confidence: 99%

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Multifunctional facets of retrovirus integrase

Grandgenett¹

2015

WJBC

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The retrovirus integrase (IN) is responsible for integration of the reverse transcribed linear cDNA into the host DNA genome. First, IN cleaves a dinucleotide from the 3' OH blunt ends of the viral DNA exposing the highly conserved CA sequence in the recessed ends. IN utilizes the 3' OH ends to catalyze the concerted integration of the two ends into opposite strands of the cellular DNA producing 4 to 6 bp staggered insertions, depending on the retrovirus species. The staggered ends are repaired by host cell machinery that results in a permanent copy of the viral DNA in the cellular genome. 83-94 ISSN 1949-8454 (online)

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Section: Solution Properties Of Inmentioning

confidence: 99%

Section: Solution Properties Of Inmentioning

confidence: 99%

Section: Solution Properties Of Inmentioning

confidence: 99%

Section: Hiv and Rsv Synaptic Complexes Are Kinetically Stabilized Bymentioning

confidence: 99%

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Multifunctional facets of retrovirus integrase

Grandgenett¹

2015

WJBC

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“…We developed a one-step direct assembly method to produce soluble SSC that allowed us to investigate the assembly mechanisms of the SSC containing either IN tetramers or octamers whose molecular mass was determined by size-exclusion chromatography-multiangle light scattering (SEC-MALS) (7,10). Additional SEC-MALS and protein-protein cross-linking analyses of the soluble RSV STC further established the presence of IN octamers (2,7).…”

mentioning

confidence: 99%

Differential assembly of Rous sarcoma virus tetrameric and octameric intasomes is regulated by the C-terminal domain and tail region of integrase

Bera

Pandey

Aihara

et al. 2018

Journal of Biological Chemistry

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Retrovirus integrase (IN) catalyzes the concerted integration of linear viral DNA ends into chromosomes. The atomic structures of five different retrovirus IN-DNA complexes, termed intasomes, have revealed varying IN subunit compositions ranging from tetramers to octamers, dodecamers, and hexadecamers. Intasomes containing two IN-associated viral DNA ends capable of concerted integration are termed stable synaptic complexes (SSC), and those formed with a viral/target DNA substrate representing the product of strand-transfer reactions are strand-transfer complexes (STC). Here, we investigated the mechanisms associated with the assembly of the Rous sarcoma virus SSC and STC. C-terminal truncations of WT IN (286 residues) indicated a role of the last 18 residues ("tail" region) in assembly of the tetrameric and octameric SSC, physically stabilized by HIV-1 IN strand-transfer inhibitors. Fine mapping through C-terminal truncations and site-directed mutagenesis suggested that at least three residues (Asp-268-Thr-270) past the last β-strand in the C-terminal domain (CTD) are necessary for assembly of the octameric SSC. In contrast, the assembly of the octameric STC was independent of the last 18 residues of IN. Single-site substitutions in the CTD affected the assembly of the SSC, but not necessarily of the STC, suggesting that STC assembly may depend less on specific interactions of the CTD with viral DNA. Additionally, we demonstrate that trans-communication between IN dimer-DNA complexes facilitates the association of native long-terminal repeat (LTR) ends with partially defective LTR ends to produce a hybrid octameric SSC. The differential assembly of the tetrameric and octameric SSC improves our understanding of intasomes.

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Oligomerization of Retrovirus Integrases

Grandgenett

Aihara

2018

Subcellular Biochemistry

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Integration of the reverse-transcribed viral cDNA into the host's genome is a critical step in the lifecycle of all retroviruses. Retrovirus integration is carried out by integrase (IN), a virus-encoded enzyme that forms an oligomeric 'intasome' complex with both ends of the linear viral DNA to catalyze their concerted insertions into the backbones of the host's DNA. IN also forms a complex with host proteins, which guides the intasome to the host's chromosome. Recent structural studies have revealed remarkable diversity as well as conserved features among the architectures of the intasome assembly from different genera of retroviruses. This chapter will review how IN oligomerizes to achieve its function, with particular focus on alpharetrovirus including the avian retrovirus Rous sarcoma virus. Another chapter (Craigie) will focus on the structure and function of IN from HIV-1.

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Rous Sarcoma Virus Synaptic Complex Capable of Concerted Integration Is Kinetically Trapped by Human Immunodeficiency Virus Integrase Strand Transfer Inhibitors

Cited by 8 publications

References 43 publications

Multifunctional facets of retrovirus integrase

Multifunctional facets of retrovirus integrase

Differential assembly of Rous sarcoma virus tetrameric and octameric intasomes is regulated by the C-terminal domain and tail region of integrase

Oligomerization of Retrovirus Integrases

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