A5071647749- HIV Research and Treatment
- interferon and immune responses
- Ubiquitin and proteasome pathways
- Viral Infections and Immunology Research
- RNA and protein synthesis mechanisms
- Cytomegalovirus and herpesvirus research
- RNA Interference and Gene Delivery
- Glycosylation and Glycoproteins Research
- Immune Cell Function and Interaction
- Virology and Viral Diseases
- Peptidase Inhibition and Analysis
- DNA Repair Mechanisms
- Advanced biosensing and bioanalysis techniques
- Genetics and Neurodevelopmental Disorders
- Virus-based gene therapy research
- Cancer-related Molecular Pathways
- Herpesvirus Infections and Treatments
- CRISPR and Genetic Engineering
- Mitochondrial Function and Pathology
- Toxoplasma gondii Research Studies
- Genetics, Bioinformatics, and Biomedical Research
- HIV/AIDS drug development and treatment
- Genomics and Chromatin Dynamics
- Influenza Virus Research Studies
University of Utah
2008-2025
University of Washington
2006-2007
Seattle University
2007
HIV-1 replication and integration in vitro To infect a host cell, must reverse transcribe its single-stranded RNA genome into double-stranded DNA copy integrate that chromosome. Reverse transcription have been characterized separately but not reconstituted together outside of the cell. Christensen et al. now report viral core particles can complete full cell-free system. The external capsid shell is required for efficient transcription, replicating loop out openings. Integration requires...
TRIM5 proteins are restriction factors that block retroviral infections by binding viral capsids and preventing reverse transcription. Capsid recognition is mediated C-terminal domains on TRIM5α (SPRY) or TRIMCyp (cyclophilin A), which interact weakly with capsids. Efficient capsid also requires the conserved N-terminal tripartite motifs (TRIM), mediate oligomerization create avidity effects. To characterize how recognize capsids, we developed methods for isolating native recombinant...
Article22 June 2015Open Access TRIM5α requires Ube2W to anchor Lys63-linked ubiquitin chains and restrict reverse transcription Adam J Fletcher MRC Centre of Medical Molecular Virology, Division Infection Immunity, University College London, UKThese authors contributed equally this work Search for more papers by author Devin E Christensen Department Biochemistry HSC Core Facilities, Utah School Medicine, Salt Lake City, UT, USAThese Chad Nelson USA Choon Ping Tan UK Torsten Schaller Paul...
Restriction factors and pattern recognition receptors are important components of intrinsic cellular defenses against viral infection. Mammalian TRIM5α proteins restriction that target the capsid cores retroviruses activate ubiquitin-dependent antiviral responses upon recognition. Here, we report crystallographic functional studies B-box 2 domain, which mediates higher-order assembly TRIM5 proteins. The can form both dimers trimers, trimers link multiple into a hexagonal net matches lattice...
The rhesus monkey intrinsic immunity factor TRIM5alpha(rh) recognizes incoming capsids from a variety of retroviruses, including human immunodeficiency virus type 1 (HIV-1) and equine infectious anemia (EIAV), inhibits the accumulation viral reverse transcripts. However, direct interactions between restricting TRIM5alpha proteins retroviral have not previously been demonstrated using pure recombinant proteins. To facilitate structural mechanistic studies restriction, we developed methods for...
TRIM5α combines distinct modes of binding into successively higher-order structures to recognize HIV-1 and retroviral capsids.
Retroviruses can be detected by the innate immune sensor cyclic GMP-AMP synthase (cGAS), which recognizes reverse-transcribed DNA and activates an antiviral response. However, extent to HIV-1 shields its genome from cGAS recognition remains unclear. To study this process in mechanistic detail, we reconstituted reverse transcription, release, sensing of a cell-free system. We found that wild-type capsids protect viral genomes even after completing transcription. Viral could “deprotected”...
Transient transfection of small interfering RNA (siRNA) provides a powerful approach for studying cellular protein functions, particularly when the target can be re-expressed from an exogenous siRNA-resistant construct in order to rescue knockdown phenotype, confirm siRNA specificity, and support mutational analyses. Rescue experiments often fail, however, constructs are expressed at suboptimal levels. Here, we describe ensemble mammalian expression vectors with CMV promoters differing...
HIV-1 DNA is preferentially integrated into chromosomal hot spots by the preintegration complex (PIC). To understand mechanism, we measured integration activity of PICs-extracted from infected cells-and intasomes, biochemically assembled PIC substructures using a number relevant target substrates. We observed that PIC-mediated human chromatin preferred compared to genomic DNA. Surprisingly, nucleosomes lacking histone modifications were not analogous naked Nucleosomes containing...
ABSTRACT Restriction factors are intrinsic cellular defense proteins that have evolved to block microbial infections. Retroviruses such as HIV-1 restricted by TRIM5 proteins, which recognize the viral capsid shell surrounds, organizes, and protects genome. TRIM5α uses a SPRY domain bind capsids with low affinity ( K D of >1 mM) therefore requires higher-order assembly into hexagonal lattice generate sufficient avidity for productive recognition. TRIMCyp, on other hand, binds through...
Retroviruses can be detected by the innate immune sensor cyclic GMP-AMP synthase (cGAS), which recognizes reverse-transcribed DNA and activates an antiviral response. However, extent to HIV-1 shields its genome from cGAS recognition remains unclear. To study this process in mechanistic detail, we reconstituted reverse transcription, release, sensing of a cell-free system. We found that wild-type capsids protect viral genomes even after completing transcription. Viral could "deprotected"...