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Title: Structural basis of HIV-1 capsid recognition by PF74 and CPSF6

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [2];  [3];  [4];  [4];  [1];  [1];  [4];  [4];  [3];  [1];  [4]
  1. Univ. of Texas Health Science Center at San Antonio, San Antonio, TX (United States)
  2. Univ. of Utah, Salt Lake City, UT (United States)
  3. Albert Einstein College of Medicine, Bronx, NY (United States)
  4. Univ. of Virginia, Charlottesville, VA (United States)

Upon infection of susceptible cells by HIV-1, the conical capsid formed by ~250 hexamers and 12 pentamers of the CA protein is delivered to the cytoplasm. In this study, the capsid shields the RNA genome and proteins required for reverse transcription. In addition, the surface of the capsid mediates numerous host–virus interactions, which either promote infection or enable viral restriction by innate immune responses. In the intact capsid, there is an intermolecular interface between the N-terminal domain (NTD) of one subunit and the C-terminal domain (CTD) of the adjacent subunit within the same hexameric ring. The NTD–CTD interface is critical for capsid assembly, both as an architectural element of the CA hexamer and pentamer and as a mechanistic element for generating lattice curvature. Here we report biochemical experiments showing that PF-3450074 (PF74), a drug that inhibits HIV-1 infection, as well as host proteins cleavage and polyadenylation specific factor 6 (CPSF6) and nucleoporin 153 kDa (NUP153), bind to the CA hexamer with at least 10-fold higher affinities compared with nonassembled CA or isolated CA domains. The crystal structure of PF74 in complex with the CA hexamer reveals that PF74 binds in a preformed pocket encompassing the NTD–CTD interface, suggesting that the principal inhibitory target of PF74 is the assembled capsid. Likewise, CPSF6 binds in the same pocket. Given that the NTD–CTD interface is a specific molecular signature of assembled hexamers in the capsid, binding of NUP153 at this site suggests that key features of capsid architecture remain intact upon delivery of the preintegration complex to the nucleus.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1168490
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, Issue 52; ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)Copyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 174 works
Citation information provided by
Web of Science

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Multiple Roles of HIV-1 Capsid during the Virus Replication Cycle journal April 2019
Viral evasion of DNA-stimulated innate immune responses journal March 2016
Nuclear landscape of HIV-1 infection and integration journal December 2016
HIV-1 assembly, release and maturation journal June 2015
HIV-1 capsid: the multifaceted key player in HIV-1 infection journal July 2015
Restriction of HIV-1 and other retroviruses by TRIM5 journal July 2019
A highly potent long-acting small-molecule HIV-1 capsid inhibitor with efficacy in a humanized mouse model journal September 2019
Quantitative monitoring of the cytoplasmic release of NCp7 proteins from individual HIV-1 viral cores during the early steps of infection journal January 2019
Discovery of novel 1,4-disubstituted 1,2,3-triazole phenylalanine derivatives as HIV-1 capsid inhibitors journal January 2019
Derivation of simian tropic HIV-1 infectious clone reveals virus adaptation to a new host journal May 2019
Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes posted_content May 2019
Predicting mutations in HIV-1 Gag: Insights from in silico and an in vitro BSL2 platform on thermostability and allosteric effects journal July 2020
Identification of Chikungunya virus nucleocapsid core assembly modulators posted_content January 2019
X-ray crystal structures of native HIV-1 capsid protein reveal conformational variability journal June 2015
Ebselen, a Small-Molecule Capsid Inhibitor of HIV-1 Replication journal January 2016
Capsid-CPSF6 Interaction Is Dispensable for HIV-1 Replication in Primary Cells but Is Selected during Virus Passage In Vivo journal June 2016
HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry journal June 2015
Complementary Assays Reveal a Low Level of CA Associated with Viral Complexes in the Nuclei of HIV-1-Infected Cells journal March 2015
K63-Linked Ubiquitin Is Required for Restriction of HIV-1 Reverse Transcription and Capsid Destabilization by Rhesus TRIM5α journal May 2019
A Novel Phenotype Links HIV-1 Capsid Stability to cGAS-Mediated DNA Sensing journal June 2019
PF74 Reinforces the HIV-1 Capsid To Impair Reverse Transcription-Induced Uncoating journal August 2018
Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid journal September 2019
PF74 Inhibits HIV-1 Integration by Altering the Composition of the Preintegration Complex journal December 2018
Inhibition of HIV-1 Maturation via Small-Molecule Targeting of the Amino-Terminal Domain in the Viral Capsid Protein journal February 2017
Analysis of CA Content and CPSF6 Dependence of Early HIV-1 Replication Complexes in SupT1-R5 Cells journal November 2019
HIV-1 capsid is involved in post-nuclear entry steps journal April 2016
Impact of LEDGIN treatment during virus production on residual HIV-1 transcription journal April 2019
MxB impedes the NUP358-mediated HIV-1 pre-integration complex nuclear import and viral replication cooperatively with CPSF6 journal June 2020
KIF5B and Nup358 Cooperatively Mediate the Nuclear Import of HIV-1 during Infection journal June 2016
Time-Resolved Imaging of Single HIV-1 Uncoating In Vitro and in Living Cells journal June 2016
A Novel Entry/Uncoating Assay Reveals the Presence of at Least Two Species of Viral Capsids During Synchronized HIV-1 Infection journal September 2016
Digoxin reveals a functional connection between HIV-1 integration preference and T-cell activation journal July 2017
GS-CA Compounds: First-In-Class HIV-1 Capsid Inhibitors Covering Multiple Grounds journal June 2019
HIV Capsid Inhibitors Beyond PF74 journal October 2019
Review of Current Cell-Penetrating Antibody Developments for HIV-1 Therapy journal February 2018
Live-Cell Imaging of Early Steps of Single HIV-1 Infection journal May 2018
Kinetics of HIV-1 capsid uncoating revealed by single-molecule analysis journal June 2018
HIV-1 nuclear import in macrophages is regulated by CPSF6-capsid interactions at the nuclear pore complex journal January 2019
Fluorescence Biosensor for Real-Time Interaction Dynamics of Host Proteins with HIV-1 Capsid Tubes journal June 2019
Defining Pharmacological Targets by Analysis of Virus–Host Protein Interactions book January 2018
Chromatin organization at the nuclear pore favours HIV replication journal March 2015
Exploring histone loading on HIV DNA reveals a dynamic nucleosome positioning between unintegrated and integrated viral genome journal March 2020
Extra-Low-Frequency Pulse Stimulated Conformational Change in Blood-Cell Proteins and Consequent Immune Activity Transformation journal January 2020
Evolutionary and Functional Analysis of Old World Primate TRIM5 Reveals the Ancient Emergence of Primate Lentiviruses and Convergent Evolution Targeting a Conserved Capsid Interface journal August 2015
Comparison of Biochemical Properties of HIV-1 and HIV-2 Capsid Proteins journal June 2017
Visualizing HIV-1 Capsid and Its Interactions with Antivirals and Host Factors journal February 2021
Insights into Adenovirus Uncoating from Interactions with Integrins and Mediators of Host Immunity journal December 2016
Closing the net on retroviruses journal July 2016