Recruitment of a SAP18-HDAC1 complex into HIV-1 virions and its requirement for viral replication

Masha Sorin, Jennifer Cano, Supratik Das, Sheeba Mathew, Xuhong Wu, Kelvin Davies, Xuanling Shi, S. W Grace Cheng, David Ott, Ganjam V. Kalpana

Research output: Contribution to journalArticle

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Abstract

HIV-1 integrase (IN) is a virally encoded protein required for integration of viral cDNA into host chromosomes. INI1/hSNF5 is a component of the SWI/SNF complex that interacts with HIV-1 IN, is selectively incorporated into HIV-1 (but not other retroviral) virions, and modulates multiple steps, including particle production and infectivity. To gain further insight into the role of INI1 in HIV-1 replication, we screened for INI1-interacting proteins using the yeast two-hybrid system. We found that SAP18 (Sin3a associated protein 18 kD), a component of the Sin3a-HDAC1 complex, directly binds to INI1 in yeast, in vitro and in vivo. Interestingly, we found that IN also binds to SAP18 in vitro and in vivo. SAP18 and components of a Sin3A-HDAC1 complex were specifically incorporated into HIV-1 (but not SIV and HTLV-1) virions in an HIV-1 IN-dependent manner. Using a fluorescence-based assay, we found that HIV-1 (but not SIV) virion preparations harbour significant deacetylase activity, indicating the specific recruitment of catalytically active HDAC into the virions. To determine the requirement of virion-associated HDAC1 to HIV-1 replication, an inactive, transdominant negative mutant of HDAC1 (HDAC1 H141A) was utilized. Incorporation of HDAC1H141A decreased the virion-associated histone deacetylase activity. Furthermore, incorporation of HDAC1H141A decreased the infectivity of HIV-1 (but not SIV) virions. The block in infectivity due to virion-associated HDAC1H141A occurred specifically at the early reverse transcription stage, while entry of the virions was unaffected. RNA-interference mediated knock-down of HDAC1 in producer cells resulted in decreased virion-associated HDAC1 activity and a reduction in infectivity of these virions. These studies indicate that HIV-1 IN and INI1/hSNF5 bind SAP18 and selectively recruit components of Sin3a-HDAC1 complex into HIV-1 virions. Furthermore, HIV-1 virion-associated HDAC1 is required for efficient early post-entry events, indicating a novel role for HDAC1 during HIV-1 replication.

Original languageEnglish (US)
Article numbere1000463
JournalPLoS Pathogens
Volume5
Issue number6
DOIs
StatePublished - Jun 2009

Fingerprint

Virion
HIV-1
Proteins
Virus Integration
Integrases
Two-Hybrid System Techniques
Human T-lymphotropic virus 1
Histone Deacetylases
RNA Interference
Reverse Transcription
Complementary DNA
Chromosomes
Yeasts
Fluorescence

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Recruitment of a SAP18-HDAC1 complex into HIV-1 virions and its requirement for viral replication. / Sorin, Masha; Cano, Jennifer; Das, Supratik; Mathew, Sheeba; Wu, Xuhong; Davies, Kelvin; Shi, Xuanling; Cheng, S. W Grace; Ott, David; Kalpana, Ganjam V.

In: PLoS Pathogens, Vol. 5, No. 6, e1000463, 06.2009.

Research output: Contribution to journalArticle

Sorin, Masha ; Cano, Jennifer ; Das, Supratik ; Mathew, Sheeba ; Wu, Xuhong ; Davies, Kelvin ; Shi, Xuanling ; Cheng, S. W Grace ; Ott, David ; Kalpana, Ganjam V. / Recruitment of a SAP18-HDAC1 complex into HIV-1 virions and its requirement for viral replication. In: PLoS Pathogens. 2009 ; Vol. 5, No. 6.
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abstract = "HIV-1 integrase (IN) is a virally encoded protein required for integration of viral cDNA into host chromosomes. INI1/hSNF5 is a component of the SWI/SNF complex that interacts with HIV-1 IN, is selectively incorporated into HIV-1 (but not other retroviral) virions, and modulates multiple steps, including particle production and infectivity. To gain further insight into the role of INI1 in HIV-1 replication, we screened for INI1-interacting proteins using the yeast two-hybrid system. We found that SAP18 (Sin3a associated protein 18 kD), a component of the Sin3a-HDAC1 complex, directly binds to INI1 in yeast, in vitro and in vivo. Interestingly, we found that IN also binds to SAP18 in vitro and in vivo. SAP18 and components of a Sin3A-HDAC1 complex were specifically incorporated into HIV-1 (but not SIV and HTLV-1) virions in an HIV-1 IN-dependent manner. Using a fluorescence-based assay, we found that HIV-1 (but not SIV) virion preparations harbour significant deacetylase activity, indicating the specific recruitment of catalytically active HDAC into the virions. To determine the requirement of virion-associated HDAC1 to HIV-1 replication, an inactive, transdominant negative mutant of HDAC1 (HDAC1 H141A) was utilized. Incorporation of HDAC1H141A decreased the virion-associated histone deacetylase activity. Furthermore, incorporation of HDAC1H141A decreased the infectivity of HIV-1 (but not SIV) virions. The block in infectivity due to virion-associated HDAC1H141A occurred specifically at the early reverse transcription stage, while entry of the virions was unaffected. RNA-interference mediated knock-down of HDAC1 in producer cells resulted in decreased virion-associated HDAC1 activity and a reduction in infectivity of these virions. These studies indicate that HIV-1 IN and INI1/hSNF5 bind SAP18 and selectively recruit components of Sin3a-HDAC1 complex into HIV-1 virions. Furthermore, HIV-1 virion-associated HDAC1 is required for efficient early post-entry events, indicating a novel role for HDAC1 during HIV-1 replication.",
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AU - Wu, Xuhong

AU - Davies, Kelvin

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AU - Cheng, S. W Grace

AU - Ott, David

AU - Kalpana, Ganjam V.

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