Abstract

P5. Abstract Retroviral replication depends on integration of reverse transcribed viral DNA into a host cell chromosome. This process is catalyzed by integrase (IN) in the context of a stable synaptic complex, known as the intasome. The intasome is the target for IN strand transfer inhibitors (INSTIs), the only class of HIV IN antagonists currently approved to treat HIV/AIDS. Recent years saw characterization of the intasomes from several retroviral species, collectively elucidating the conserved intasomal core assembly (CIC) responsible for integration. We have now established the first lentiviral intasome model based on maedi-visna virus (MVV), which affords detailed structural studies of lentiviral integration in the absence of solubilizing or hyperactivating mutations in IN. The preliminary study of the MVV intasome by single particle cryo-EM in the Cherepanov laboratory revealed that it is much larger than its non-lentiviral counterparts, comprising a hexadecamer (tetramer-of-tetramers) of IN. In the proposed project we take advantage of the new system to study lentiviral DNA integration into chromatin and the role of the host cell factor LEDGF/p75 in this process. As a translational component of this project, we will develop a lentiviral intasome model for structural studies of INSTIs and the mechanisms of drug resistance.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center (P50)
Project #
2P50GM082251-11
Application #
9407943
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2017-08-10
Budget End
2018-07-31
Support Year
11
Fiscal Year
2017
Total Cost
Indirect Cost

  Institution

Name
University of Pittsburgh
Department
Type
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213

  Publications

Martin, Jessica L; Mendonça, Luiza M; Marusinec, Rachel et al. (2018) Critical Role of the Human T-Cell Leukemia Virus Type 1 Capsid N-Terminal Domain for Gag-Gag Interactions and Virus Particle Assembly. J Virol 92:
Wang, Mingzhang; Lu, Manman; Fritz, Matthew P et al. (2018) Fast Magic-Angle Spinning 19 F?NMR Spectroscopy of HIV-1 Capsid Protein Assemblies. Angew Chem Int Ed Engl 57:16375-16379
Paramasivam, Sivakumar; Gronenborn, Angela M; Polenova, Tatyana (2018) Backbone amide 15N chemical shift tensors report on hydrogen bonding interactions in proteins: A magic angle spinning NMR study. Solid State Nucl Magn Reson 92:1-6
Fritz, Matthew; Quinn, Caitlin M; Wang, Mingzhang et al. (2018) Determination of accurate backbone chemical shift tensors in microcrystalline proteins by integrating MAS NMR and QM/MM. Phys Chem Chem Phys 20:9543-9553
Quinn, Caitlin M; Wang, Mingzhang; Fritz, Matthew P et al. (2018) Dynamic regulation of HIV-1 capsid interaction with the restriction factor TRIM5? identified by magic-angle spinning NMR and molecular dynamics simulations. Proc Natl Acad Sci U S A 115:11519-11524
Varlakhanova, Natalia V; Alvarez, Frances J D; Brady, Tyler M et al. (2018) Structures of the fungal dynamin-related protein Vps1 reveal a unique, open helical architecture. J Cell Biol 217:3608-3624
Ning, Jiying; Zhong, Zhou; Fischer, Douglas K et al. (2018) Truncated CPSF6 Forms Higher-Order Complexes That Bind and Disrupt HIV-1 Capsid. J Virol 92:
Himes, Benjamin A; Zhang, Peijun (2018) emClarity: software for high-resolution cryo-electron tomography and subtomogram averaging. Nat Methods 15:955-961
Balasubramaniam, Muthukumar; Zhou, Jing; Addai, Amma et al. (2018) PF74 Inhibits HIV-1 Integration by Altering The Composition of the Preintegration Complex. J Virol :
Lu, Manman; Sarkar, Sucharita; Wang, Mingzhang et al. (2018) 19F Magic Angle Spinning NMR Spectroscopy and Density Functional Theory Calculations of Fluorosubstituted Tryptophans: Integrating Experiment and Theory for Accurate Determination of Chemical Shift Tensors. J Phys Chem B 122:6148-6155

Showing the most recent 10 out of 144 publications