A5058005825- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- Biochemical and Molecular Research
- Cytomegalovirus and herpesvirus research
- CRISPR and Genetic Engineering
- RNA Research and Splicing
- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- HIV/AIDS Research and Interventions
- Nuclear Structure and Function
- RNA modifications and cancer
- Immune Cell Function and Interaction
- Bacteriophages and microbial interactions
- Herpesvirus Infections and Treatments
- Signaling Pathways in Disease
- RNA and protein synthesis mechanisms
- Genomics and Chromatin Dynamics
- Plant Virus Research Studies
- DNA and Nucleic Acid Chemistry
- interferon and immune responses
- Monoclonal and Polyclonal Antibodies Research
- Advanced biosensing and bioanalysis techniques
- Ubiquitin and proteasome pathways
- Pluripotent Stem Cells Research
- Chronic Lymphocytic Leukemia Research
Dana-Farber Cancer Institute
2016-2025
Harvard University
2016-2025
University of Pittsburgh
2017-2025
Harvard University Press
2012-2021
Boston University
1988-2019
Creative Research Enterprises (United States)
2017
Technische Universität Dresden
2016
Chromosome 18 Registry & Research Society
2016
Institute for Molecular Medicine
2016
University Hospital Carl Gustav Carus
2016
HIV-1 exploits multiple host proteins during infection. We performed a large-scale small interfering RNA screen to identify factors required by and identified more than 250 HIV-dependency (HDFs). These participate in broad array of cellular functions implicate new pathways the viral life cycle. Further analysis revealed previously unknown roles for retrograde Golgi transport (Rab6 Vps53) entry, karyopherin (TNPO3) integration, Mediator complex (Med28) transcription. Transcriptional that HDF...
HIV integrase is the enzyme responsible for inserting viral DNA into host chromosome; it essential replication. The crystal structure of catalytically active core domain (residues 50 to 212) HIV-1 was determined at 2.5 Å resolution. central feature a five-stranded β sheet flanked by helical regions. overall topology reveals that this belongs superfamily polynucleotidyl transferases includes ribonuclease H and Holliday junction resolvase RuvC. site region identified position two conserved...
We have probed the structural organization of human immunodeficiency virus type 1 integrase protein by limited proteolysis and functional site-directed mutagenesis selected amino acid residues. A central region was relatively resistant to proteolysis. Proteins with altered acids in this region, or N-terminal part that includes a putative zinc-binding motif, were purified assayed for 3' processing, DNA strand transfer, disintegration activities vitro. In general, these mutations had parallel...
LEDGF/p75 directly interacts with lentiviral integrase proteins and can modulate their enzymatic activities chromosomal association. A novel genetic knockout model was established that allowed us for the first time to analyze HIV-1 integration in absence of protein. Supporting a crucial role cofactor viral replication, vector reporter gene expression were significantly reduced LEDGF-null cells. Yet, processed cDNA termini normally maintained its local target DNA sequence preference during...
The integration of a DNA copy the human immunodeficiency virus type 1 (HIV-1) genome into chromosome an infected cell is pivotal step in replication. Integration requires activity virus-encoded integrase, which enters as component virion. Results numerous mutagenesis studies have identified amino acid residues and protein domains HIV-1 integrase critical for vitro activity, but only few these mutants been studied their effects on HIV We introduced site-directed changes infectious clone show...
Integrase (IN) is an essential retroviral enzyme, and human transcriptional coactivator p75, which also referred to as lens epithelium-derived growth factor (LEDGF), the dominant cellular binding partner of HIV-1 IN. Here, we report crystal structure dimeric catalytic core domain IN complexed IN-binding LEDGF. Previously identified LEDGF hotspot residues anchor protein both monomers at dimer interface. The principal structural features that are recognized by host backbone conformation...
Lentiviruses can infect non-dividing cells, and various cellular transport proteins provide crucial functions for lentiviral nuclear entry integration. We previously showed that the viral capsid (CA) protein mediated dependency on nucleoporin (NUP) 153 during HIV-1 infection, now demonstrate a direct interaction between CA N-terminal domain phenylalanine-glycine (FG)-repeat enriched NUP153 C-terminal (NUP153C). NUP153C fused to effector domains of rhesus Trim5α restriction factor...
Significance HIV-1 requires integration for efficient gene expression, and the local chromatin environment significantly influences level of transcription. Silent, integrated proviruses constitute latent HIV reservoir. As commandeers cellular factors to dictate its preferred sites, these interactions consequentially influence latency. We examined impact polyadenylation specificity factor CPSF6, which binds capsid, integrase-binding reader LEDGF/p75 on viral infection site distribution....
The potent HIV-1 capsid inhibitor GS-6207 is an investigational principal component of long-acting antiretroviral therapy. We found that inhibits by stabilizing and thereby preventing functional disassembly the shell in infected cells. X-ray crystallography, cryo-electron microscopy, hydrogen-deuterium exchange experiments revealed tightly binds two adjoining subunits promotes distal intra- inter-hexamer interactions stabilize curved lattice. In addition, interferes with binding to cellular...
The integrase protein of human immunodeficiency virus type 1 carries out a set polynucleotidyl transfer reactions that result in the covalent attachment retroviral cDNA to host DNA. We have analyzed activities deletion derivatives protein. analysis reveals central domain only 137 amino acids is sufficient vitro catalyze subset carried by complete This polypeptide contains an acid sequence motif, Asp-Xaa39-58-Asp-Xaa35-Glu (DX39-58DX35E, where X and subscript indicate intervening between...
The solution structure of the DNA binding domain HIV-1 integrase (residues 220-270) has been determined by multidimensional NMR spectroscopy. protein is a dimer in solution, and each subunit composed five-stranded beta-barrel with topology very similar to that SH3 domain. formed stacked beta-interface comprising strands 2, 3, 4, two triple-stranded antiparallel beta-sheets, one from subunit, oriented other. One surface dimer, bounded loop between beta 1 forms saddle-shaped groove dimensions...
Structural studies of human immunodeficiency virus type 1 (HIV-1) integrase have been impeded by the low solubility protein. By systematic replacement hydrophobic residues, we previously identified a single amino acid change (F185K) that dramatically improved catalytic domain HIV-1 and enabled structure to be determined x-ray crystallography. We introduced same mutation into full-length integrase. The resulting recombinant protein is soluble fully active in vitro, whereas, carrying...
Human lens epithelium-derived growth factor/transcriptional co-activator p75 (LEDGF/p75) protein was recently identified as a binding partner for HIV-1 integrase (IN) in human cells. In this work, we used biochemical and bioinformatic approaches to define the domain organization of LEDGF/p75. Using limited proteolysis deletion mutagenesis show that contains pair evolutionarily conserved domains, assuming about 35% its sequence. Whereas N-terminal PWWP had been recognized previously, second...