A5071154330- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- Biochemical and Molecular Research
- Cancer-related gene regulation
- RNA and protein synthesis mechanisms
- Advanced Electron Microscopy Techniques and Applications
- RNA modifications and cancer
- Electron and X-Ray Spectroscopy Techniques
- Enzyme Structure and Function
- RNA Research and Splicing
- Epigenetics and DNA Methylation
- Protein Kinase Regulation and GTPase Signaling
- Protein Structure and Dynamics
- Computational Drug Discovery Methods
- Fungal and yeast genetics research
- Ubiquitin and proteasome pathways
- RNA regulation and disease
- NF-κB Signaling Pathways
- Monoclonal and Polyclonal Antibodies Research
- Neuroscience and Neuropharmacology Research
- Anodic Oxide Films and Nanostructures
- Cancer-related Molecular Pathways
- Cytokine Signaling Pathways and Interactions
- Genomics and Chromatin Dynamics
- HIV-related health complications and treatments
Salk Institute for Biological Studies
2015-2024
Scripps Research Institute
2014
Universidade de São Paulo
2008-2012
Howard Hughes Medical Institute
2008-2009
Brazilian Synchrotron Light Laboratory
2002-2009
Universidade Estadual de Campinas (UNICAMP)
2002-2009
University of Arizona
2008
Like all retroviruses, HIV-1 irreversibly inserts a viral DNA (vDNA) copy of its RNA genome into host target (tDNA). The intasome, higher-order nucleoprotein complex composed integrase (IN) and the ends linear vDNA, mediates integration. Productive integration chromatin results in formation strand transfer (STC) containing catalytically joined vDNA tDNA. intasomes have been refractory to high-resolution structural studies. We used soluble IN fusion protein facilitate studies, through which...
Significance All organisms have systems in place to ensure that aberrant nascent polypeptide chains are promptly dealt with before being released from ribosomes and posing harm the cell. The ribosome-associated quality control complex (RQC), composed of Ltn1 E3 ubiquitin ligase catalytic subunit cofactors, has become a paradigm for understanding eukaryotes. However, exactly how RQC functions remained unknown. Here, we determine structure 60S subunit-bound complex. data provide critical...
Strengths and weaknesses of an HIV drug Retroviruses replicate by inserting a copy their RNA, which has been reverse transcribed into DNA, the host genome. This process involves intasome, nucleoprotein complex comprising copies viral integrase bound at ends DNA. strand-transfer inhibitors (INSTIs) stop from replicating blocking are widely used in treatment. Cook et al. describe structures second-generation to simian immunodeficiency virus (SIV) intasome with mutations known cause resistance....
Abstract Structural biology efforts using cryogenic electron microscopy are frequently stifled by specimens adopting “preferred orientations” on grids, leading to anisotropic map resolution and impeding structure determination. Tilting the specimen stage during data collection is a generalizable solution but has historically led substantial attenuation. Here, we develop updated image processing workflows demonstrate, multiple specimens, that attenuation negligible or significantly reduced...
Eukaryotic mRNAs are subject to quality control mechanisms that degrade defective mRNAs. In yeast, with stalls in translation elongation targeted for endonucleolytic cleavage by No-Go decay (NGD). The triggered is dependent on Dom34p and Hbs1p, Dom34 has been proposed be the endonuclease responsible mRNA cleavage. We created several mutants examined their effects NGD yeast. identified mutations loops of structure affect NGD. contrast, inactivating nuclease domain do not vivo. Moreover, we...
HIV-1 infection depends on the integration of viral DNA into host chromatin. Integration is mediated by enzyme integrase and blocked strand transfer inhibitors (INSTIs), first-line antiretroviral therapeutics widely used in clinic. Resistance to even best INSTIs a problem, mechanisms resistance are poorly understood. Here, we analyze combinations mutations E138K, G140A/S, Q148H/K/R, which confer INSTIs. The investigational drug 4d more effectively inhibited mutants compared with approved...
Distinct signaling pathways activate the NF-κB family of transcription factors. The canonical NF-κB-signaling pathway is mediated by IκB kinase 2/β (IKK2/β), while non-canonical depends on IKK1/α. structural and biochemical bases for distinct these otherwise highly similar IKKs are unclear. We report single-particle cryoelectron microscopy (cryo-EM) X-ray crystal structures human IKK1 in dimeric (∼150 kDa) hexameric (∼450 forms. hexamer, which representative form but comprises only ∼2%...
The currently recommended first-line therapy for HIV-1-infected patients is an integrase (IN) strand transfer inhibitor (INSTI), either dolutegravir (DTG) or bictegravir (BIC), in combination with two nucleoside reverse transcriptase inhibitors (NRTIs). Both DTG and BIC potently inhibit most INSTI-resistant IN mutants selected by the INSTIs raltegravir (RAL) elvitegravir (EVG). has not been reported to select resistance treatment-naive patients, a small number of resistant viruses patients....
Key proteins of retroviruses and other RNA viruses are translated subsequently processed from polyprotein precursors by the viral protease (PR). Processing HIV Gag-Pol yields structural enzymes. Structures mature enzymes PR, reverse transcriptase (RT), integrase (IN) aided understanding catalysis design antiretrovirals, but knowledge Pol precursor architecture function before PR cleavage is limited. We developed a system to produce stable HIV-1 determined its cryo–electron microscopy...
The two human proteins Ki-1/57 and CGI-55 have highly similar amino acid sequences but their functions are unknown. We analyzed them by yeast two-hybrid screens found that they interact with the C-terminal region of chromatin-remodeling factor CHD-3 (chromo-helicase-DNA-binding domain protein-3). interaction could be confirmed in vitro vivo co-immunoprecipitations from Sf9 insect cells. Mapping showed interacts via regions at its N- C-terminals. mRNAs show highest expression muscle, colon...
The human 57 kDa Ki‐1 antigen (Ki‐1/57) is a cytoplasmic and nuclear protein, associated with Ser/Thr protein kinase activity, phosphorylated at the serine threonine residues upon cellular activation. We have shown that Ki‐1/57 interacts chromo‐helicase DNA‐binding domain 3 adaptor/signaling receptor of activated 1 in nucleus. Among identified proteins interacted yeast two‐hybrid system was arginine‐methyltransferase‐1 (PRMT1). Most interestingly, when PRMT1 used as bait we were able to...
Ki-1/57, the 57-kDa human protein antigen recognized by CD30 antibody Ki-1, is a cytoplasmic and nuclear that phosphorylated on serine threonine residues. When isolated from Hodgkin's lymphoma analogous cell line L540 Ki-1/57 co-immunoprecipitated with Thr/Ser kinase activity. It has been also found to interact hyaluronic acid therefore termed intracellular IHABP4 (hyaluronan-binding 4). Recent studies demonstrated, however, engages in specific interaction chromo-helicase-DNA-binding domain...
Plasmodium falciparum causes the most severe form of malaria and is responsible for majority deaths worldwide. The mechanism cell cycle control within intra-erythrocytic stages has been examined as a potential means promising way to identifying how stop parasite development in red blood cells. Our group determined that melatonin increases parasitemia P. chabaudi through complex signalling cascade. In vertebrates, controls expression transcription factors, leading us postulate rather...
Integrase strand transfer inhibitors (INSTIs) block the integration step of retroviral lifecycle and are first-line drugs used for treatment HIV-1/AIDS. INSTIs have a polycyclic core with heteroatom triads, chelate metal ions at active site, halobenzyl group that interacts viral DNA attached to by flexible linker. The most broadly effective inhibit both wild-type (WT) integrase (IN) variety well-known mutants. However, because there mutations reduce potency all available INSTIs, new better...
The human protein Ki-1/57 was first identified through the cross reactivity of anti-CD30 monoclonal antibody Ki-1, in Hodgkin lymphoma cells. expression diverse cancer cells and its phosphorylation peripheral blood leukocytes after mitogenic activation suggested possible role cell signaling. interacts with several other regulatory proteins involved cellular signaling, transcriptional regulation RNA metabolism, suggesting it may have pleiotropic functions. In a previous spectroscopic...
Abstract The human SFRS9/SRp30c belongs to the SR family of splicing regulators. Despite evidence that members this protein may be targeted by arginine methylation, has yet experimentally addressed. In study, we found SFRS9 is a target for PRMT1-mediated methylation in vitro, and it immunoprecipitated from HEK-293 lysates antibodies recognize both mono- dimethylated arginines. We further observed upon treatment with inhibitor Adox, fluorescent EGFP-SFRS9 re-localizes dot-like structures cell...
The cytoplasmic and nuclear protein Ki-1/57 was first identified in malignant cells from Hodgkin's lymphoma. Despite studies showing its phosphorylation, arginine methylation, interaction with several regulatory proteins, the functional role of human remains to be determined. Here, we investigated relationship RNA functions. Through immunoprecipitation assays, verified association endogenous splicing proteins hnRNPQ SFRS9 HeLa cell extracts. We also found that recombinant able bind a poly-U...
Ki-1/57 is a 57-kDa cytoplasmic and nuclear protein associated with kinase activity hyper-phosphorylated on Ser/Thr residues upon cellular activation. In previous studies we identified the receptor of activated kinase-1 (RACK1), signaling adaptor that binds PKC, as interacts Ki-1/57. Here demonstrate far-UV circular dichroism spectrum WD repeat-containing RACK1 shows an unusual positive ellipticity at 229 nm, which in other proteins family has been attributed to surface tryptophans are...