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Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes
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CPSF6 promotes HIV-1 preintegration complex function.

Evan Chaudhuri1,2,3,4, Sooin Jang5,6, Rajasree Chakraborty1,4

  • 1Center for AIDS Health Disparities Research, Nashville, Tennessee, USA.

Journal of Virology
|April 9, 2025
PubMed
Summary
This summary is machine-generated.

Cleavage and polyadenylation specificity factor 6 (CPSF6) directly enhances HIV-1 preintegration complex (PIC) activity and promotes viral DNA integration. Disrupting the CPSF6-HIV-1 capsid interaction reduces integration and redirects it away from gene-rich areas.

Keywords:
capsidcleavage and polyadenylation specificity factor 6 (CPSF6)human immunodeficiency virus (HIV)integrationpreintegration complex

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Cleavage and polyadenylation specificity factor 6 (CPSF6) is a host factor involved in mRNA processing and HIV-1 replication.
  • CPSF6 binds to the HIV-1 capsid (CA) and influences viral DNA integration targeting into gene-dense genomic regions.
  • The precise role of CPSF6 in the function of the HIV-1 preintegration complex (PIC) remained largely uncharacterized.

Purpose of the Study:

  • To investigate the effect of CPSF6 on the activity of the HIV-1 preintegration complex (PIC).
  • To determine the role of the CPSF6-CA interaction in HIV-1 DNA integration and targeting.
  • To elucidate how CPSF6 influences HIV-1 integration site selection within the host genome.

Main Methods:

  • Extraction of PICs from CPSF6-depleted or CA-binding mutant cells for in vitro integration assays.
  • Reconstitution experiments using purified recombinant CPSF6.
  • Infection of CPSF6-mutant cells with HIV-1 and measurement of viral DNA integration, reverse transcription, and nuclear entry.
  • Sequencing analysis of HIV-1 integration sites in CPSF6-mutant versus control cells.

Main Results:

  • PICs from CPSF6-depleted or CA-binding mutant cells showed significantly reduced viral DNA integration activity.
  • Addition of recombinant CPSF6 restored integration activity in mutant PICs, indicating a direct role.
  • HIV-1 integration was significantly reduced in CPSF6-mutant cells, independent of reverse transcription or nuclear entry.
  • Disruption of CPSF6-CA binding redirected HIV-1 integration away from gene-dense regions to gene-poor areas.

Conclusions:

  • The CPSF6-CA interaction is crucial for stimulating HIV-1 preintegration complex (PIC) function.
  • CPSF6 plays a direct role in promoting efficient viral DNA integration.
  • The CPSF6-CA interaction dictates HIV-1 integration site selection, favoring gene-dense genomic regions.