A5084222887- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- CRISPR and Genetic Engineering
- Cytomegalovirus and herpesvirus research
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Immune Cell Function and Interaction
- HIV/AIDS Research and Interventions
- Virus-based gene therapy research
- Herpesvirus Infections and Treatments
- Bacteriophages and microbial interactions
- Biochemical and Molecular Research
- RNA Interference and Gene Delivery
- RNA modifications and cancer
- T-cell and Retrovirus Studies
- Viral gastroenteritis research and epidemiology
- Viral Infections and Immunology Research
- Fibromyalgia and Chronic Fatigue Syndrome Research
- Nuclear Structure and Function
- RNA regulation and disease
- Digital Imaging for Blood Diseases
- Hepatitis C virus research
- Monoclonal and Polyclonal Antibodies Research
- Immunotherapy and Immune Responses
- Chromosomal and Genetic Variations
National Cancer Institute
2016-2025
Center for Cancer Research
2016-2025
Frederick National Laboratory for Cancer Research
2012-2024
Chhatrapati Shahu Ji Maharaj University
2022-2023
Dr. A.P.J. Abdul Kalam Technical University
2020-2021
Jawaharlal Nehru University
2020
Cancer Institute (WIA)
2019
Palacký University Olomouc
2017
National Institute of Standards and Technology
2013
Rice University
2013
Phosphorylation of the alpha subunit eucaryotic translation initiation factor (eIF-2 alpha) by double-stranded RNA-activated inhibitor (DAI) kinase correlates with inhibition initiation. The importance eIF-2 phosphorylation in regulating was studied expression specific mutants COS-1 cells. DNA transfection certain plasmids could activate DAI and result poor plasmid-derived mRNAs. In these cases, mRNAs improved presence inhibitors or a nonphosphorylatable mutant (serine to alanine) alpha....
Significance For several decades, retroviral core uncoating has been thought to occur in the cytoplasm coordination with reverse transcription, and while some recent studies have concluded that HIV-1 occurs at nuclear envelope during import, none nucleus. Here, we developed methods study by direct labeling quantification of viral capsid protein associated infectious cores produced transcriptionally active proviruses. We find nuclei infected cells are largely intact uncoat near their...
The retrovirus XMRV (xenotropic murine leukemia virus-related virus) has been detected in human prostate tumors and blood samples from patients with chronic fatigue syndrome, but these findings have not replicated. We hypothesized that an understanding of when how first arose might help explain the discrepant results. studied cancer cell lines CWR22Rv1 CWR-R1, which produce virtually identical to viruses recently found patient samples, as well their progenitor tumor xenograft (CWR22) had...
Significance Here, we used a fluorescent protein that is free in solution and trapped nuclear HIV-1 capsids to demonstrate the retain integrity prevent mixing of macromolecules within viral core cellular environment until just before integration. We also found capsid maintained minutes disassembly nucleus, revealing uncoating proceeds rapidly after loss. These valuable insights into early stage replication indicate intact are imported through pores, reverse transcription mostly completed...
We determined the in vivo forward mutation rate a single replication cycle for spleen necrosis virus (SNV). A method was developed to clone integrated proviruses of retroviral shuttle vectors by exploiting tight binding lac operator repressor protein. The contained lacZ alpha gene as reporter mutations. Thirty-seven 16,867 recovered five classes mutations, including substitutions and frameshifts. Runs 9 10 identical base pairs direct repeat 110 were mutational hotspots. In addition, two...
ABSTRACT Encapsidation of host restriction factor APOBEC3G (A3G) into vif -deficient human immunodeficiency virus type 1 (HIV-1) blocks replication at least partly by C-to-U deamination viral minus-strand DNA, resulting in G-to-A hypermutation. A3G may also inhibit HIV-1 reducing DNA synthesis and inducing degradation. To gain further insight the mechanisms inhibition, we examined metabolism A3G-exposed DNA. We observed that an overall 35-fold decrease infectivity was accompanied a five- to...
Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) is a host cytidine deaminase that packaged into virions and confers resistance to retroviral infection. APOBEC3G deaminates deoxycytidines in minus strand DNA deoxyuridines, resulting G A hypermutation viral inactivation. Human immunodeficiency virus type 1 (HIV-1) virion infectivity factor counteracts the antiviral activity of by inducing its proteosomal degradation preventing incorporation. To elucidate mechanism...
HIV-1 and other retroviruses occasionally undergo hypermutation, characterized by a high rate of G-to-A substitution. Recently, the human apolipoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like 3G (APOBEC3G), first identified as CEM15, was shown to be packaged into retroviral virions deaminate deoxycytidine deoxyuridine in newly synthesized viral minus-strand DNA, thereby inducing hypermutation. This innate mechanism resistance infection is counteracted infectivity factor (Vif),...
Guidelines for Naming Nonprimate APOBEC3 Genes and Proteins Rebecca S. LaRue, Valgerdur Andresdottir, Yannick Blanchard, Silvestro G. Conticello, David Derse, Michael Emerman, Warner C. Greene, Stefan R. Jonsson, Nathaniel Landau, Martin Lochelt, Harmit Malik, H. Malim, Carsten Munk, Stephen J. O’Brien, Vinay K. Pathak, Klaus Strebel, Simon Wain-Hobson, Xiao-Fang Yu, Naoya Yuhki, Reuben Harris*
Human cytidine deaminases APOBEC3G (A3G) and APOBEC3F (A3F) inhibit replication of Vif-deficient human immunodeficiency virus type 1 (HIV-1). HIV-1 Vif overcomes these host restriction factors by binding to them inducing their proteasomal degradation. The Vif-A3G Vif-A3F interactions are attractive targets for antiviral drug development because inhibiting the could allow defense mechanism control replication. It was recently reported that amino acids D(14)RMR(17) important functional...
In the preceding paper we described an experiment that determined in vivo forward mutation rate a single replication cycle for spleen necrosis virus. addition to substitutions, frameshifts, and hypermutations, mutated proviruses contained two classes of deletions. One class deletions short direct repeats at deletion junctions. Another had stretches sequences inserted this report, describe mutations, present models their generation. Detailed analysis with insertions indicates these mutations...
A long-standing question in retrovirus biology is how RNA genomes are distributed among virions. In the studies presented this report, we addressed issue by directly examining HIV-1 RNAs virions using a modified genome that contained recognition sites for BglG, an antitermination protein Escherichia coli bgl operon, which was coexpressed with fragment of BglG binding fused to fluorescent protein. Our results demonstrate majority (>90%) contain viral RNAs. We also containing or bacteriophage...
Phosphorylation of the alpha subunit eucaryotic translation initiation factor (eIF-2 alpha) by double-stranded RNA-activated inhibitor (DAI) kinase correlates with inhibition initiation. The importance eIF-2 phosphorylation in regulating was studied expression specific mutants COS-1 cells. DNA transfection certain plasmids could activate DAI and result poor plasmid-derived mRNAs. In these cases, mRNAs improved presence inhibitors or a nonphosphorylatable mutant (serine to alanine) alpha....
ABSTRACT Recent studies have shown that APOBEC3G (A3G), a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication, is localized to cytoplasmic mRNA-processing bodies (P bodies). However, the functional relevance A3G colocalization with P body marker proteins has not been established. To explore relationship between HIV-1, A3G, and bodies, we analyzed effects overexpression Mov10, DCP1a, DCP2 on HIV-1 replication. Our results show putative RNA helicase was previously...
The role of APOBEC3 (A3) protein family members in inhibiting retrovirus infection and mobile element retrotransposition is well established. However, the evolutionary effects these restriction factors may have had on active retroviruses such as HIV-1 are less understood. An variant that has been highly G-to-A mutated unlikely to be transmitted due accumulation deleterious mutations. hA3G target sequences within which mutations least more likely survive selection pressure. Thus, among...
The dynamics and regulation of HIV-1 nuclear import its intranuclear movements after have not been studied. To elucidate these essential post-entry events, we labeled viral complexes with two fluorescently tagged virion-incorporated proteins (APOBEC3F or integrase), analyzed the envelope (NE) docking, import, in living cells. We observed that exhibit unusually long NE residence times (1.5±1.6 hrs) compared to most cellular cargos, which are imported into nuclei within milliseconds....
APOBEC3 proteins inhibit HIV-1 replication in experimental systems and induce hypermutation infected patients; however, the relative contributions of several to restriction absence viral Vif protein human primary CD4(+) T cells macrophages are unknown. We observed significant inhibition HIV-1Δvif produced 293T presence APOBEC3DE (A3DE), APOBEC3F (A3F), APOBEC3G (A3G), APOBEC3H haplotype II (A3H HapII) but not APOBEC3B (A3B), APOBEC3C (A3C), or I HapI). Our previous studies showed that amino...
Increasing evidence has suggested that the HIV-1 capsid enters nucleus in a largely assembled, intact form. However, not much is known about how cone-shaped interacts with nucleoporins (NUPs) nuclear pore for crossing complex. Here, we elucidate NUP153 binds by engaging assembled protein (CA) lattice. A bipartite motif containing both canonical and noncanonical interaction modules was identified at C-terminal tail region of NUP153. The cargo-targeting phenylalanine-glycine (FG) engaged CA...
HIV-1 cores, which contain the viral genome and replication machinery, must disassemble (uncoat) during replication. However, host factors that trigger uncoating remain unidentified. Recent studies show infectious cores enter nucleus uncoat near site of integration. Here, we efficient nuclear requires synthesis a double-stranded DNA (dsDNA) >3.5 kb efficiency correlates with size. Core disruption by capsid inhibitors releases DNA, some integrates. most is degraded, indicating intact core...
Lenacapavir (GS-6207; LEN) is a potent HIV-1 capsid inhibitor approved for treating multidrug-resistant infection. LEN binds to hydrophobic pocket between neighboring (CA) proteins in hexamers and stabilizes the lattice, but its effect on capsids not fully understood. Here, we labeled with green fluorescent protein fused CA (GFP-CA) or fluid-phase GFP content marker (cmGFP) assess LEN’s impact capsids. cores GFP-CA, cmGFP, could be immunostained an anti-GFP antibody were less sensitive...
The 4-aryl-2-hydroxy-4-oxo-2-butenoic acids and their isosteric tetrazoles are among an emerging class of aryl beta-diketo (ADK)-based agents which exhibit potent inhibition HIV-1 integrase (IN)-catalyzed strand transfer (ST) processes, while having much reduced potencies against 3'-processing (3'-P) reactions. In the current study, L-708,906 (10e) 5CITEP (13b), two examples ADK inhibitors that have been reported by Merck Shionogi pharmaceutical companies, served as model leads. Structural...
We previously proposed that a balance between nucleotide excision and template RNA degradation plays an important role in nucleoside reverse transcriptase inhibitor (NRTI) resistance. To explore the predictions of this concept, we analyzed patient-derived C-terminal domains HIV-1 (RT) NRTI found when polymerase domain contained described thymidine analog resistance mutations, mutations connection increased to 3′-azido-3′-deoxythymidine (AZT) from 11-fold as much 536-fold over wild-type RT....
Understanding the mechanisms of HIV-1 drug resistance is critical for developing more effective antiretroviral agents and therapies. Based on our previously described dynamic copy-choice mechanism retroviral recombination observations that nucleoside reverse transcriptase inhibitors (NRTIs) increase frequency template switching, we propose an equilibrium exists between ( i ) NRTI incorporation, excision, resumption DNA synthesis ii degradation RNA by RNase H activity, leading to dissociation...