A5036747077- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- RNA Research and Splicing
- Nuclear Structure and Function
- Bacteriophages and microbial interactions
- Cytomegalovirus and herpesvirus research
- Botany and Plant Ecology Studies
- Polyomavirus and related diseases
- Virus-based gene therapy research
- Plant Virus Research Studies
- Genomics and Chromatin Dynamics
- Geology and Paleoclimatology Research
- Mediterranean and Iberian flora and fauna
- Plant Taxonomy and Phylogenetics
- Herpesvirus Infections and Treatments
- SARS-CoV-2 and COVID-19 Research
- Animal Virus Infections Studies
- Cell Image Analysis Techniques
- Cancer Research and Treatments
- Tree-ring climate responses
- Liver physiology and pathology
- Atomic and Subatomic Physics Research
- Cancer Cells and Metastasis
- Lipid Membrane Structure and Behavior
- Caveolin-1 and cellular processes
Heidelberg University
2016-2025
University Hospital Heidelberg
2016-2024
Medical Research Council
2021
MRC Laboratory of Molecular Biology
2021
Tesla Blatna (Czechia)
2020
Charles University
2003-2015
Czech Academy of Sciences, Institute of Molecular Genetics
2006
Human immunodeficiency virus (HIV-1) remains a major health threat. Viral capsid uncoating and nuclear import of the viral genome are critical for productive infection. The size HIV-1 is generally believed to exceed diameter pore complex (NPC), indicating that has occur prior import. Here, we combined correlative light electron microscopy with subtomogram averaging capture structural status reverse transcription-competent complexes in infected T cells. We demonstrated NPC cellulo sufficient...
HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization, and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed cDNA molecules their association with cellular proteins using fluorescence correlative-light-and-electron-microscopy (CLEM). specifically detected nuclei, not in cytoplasm. Nuclear initially co-localized fluorescent integrase fusion (IN-FP) CA (capsid) protein, cDNA-punctae separated from...
Structural changes in HIV maturation Nascent particles assemble at the plasma membrane of an infected cell and bud into a membrane-enveloped, immature virion. Assembly budding are driven by polyprotein called Gag, which consists matrix domain (MA) that is recruited to membrane, capsid (CA) responsible for self-assembly, nucleocapsid (NC) recruits viral RNA genome. Gag cleavage results structural rearrangement produces mature Qu et al . imaged electron tomography focused on MA (see...
Summary Upon infection, human immunodeficiency virus (HIV-1) releases its cone-shaped capsid into the cytoplasm of infected T-cells and macrophages. As largest known cargo, enters nuclear pore complex (NPC), driven by interactions with numerous FG-repeat nucleoporins (FG-Nups). Whether NPCs structurally adapt to passage whether capsids are modified during remains unknown, however. Here, we combined super-resolution correlative microscopy cryo electron tomography molecular simulations study...
Abstract Nuclear pore complexes (NPCs) constitute giant channels within the nuclear envelope that mediate nucleocytoplasmic exchange. NPC diameter is thought to be regulated by tension, but how such changes are physiologically linked cell differentiation, where mechanical properties of nuclei remodeled and mechanosensing occurs, remains unstudied. Here we used cryo-electron tomography show NPCs dilate during differentiation mouse embryonic stem cells into neural progenitors. In...
Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions are surrounded by a lipid bilayer from which spike (S) protein trimers protrude. Heavily glycosylated S bind the ACE2 receptor and mediate entry of into target cells. exhibits extensive conformational flexibility: it modulates exposure its binding site later undergoes complete structural rearrangement to drive fusion viral cellular membranes. The structures conformations soluble, overexpressed, purified proteins...
Abstract Nuclear pore complexes (NPCs) mediate nucleocytoplasmic exchange, which is essential for eukaryotes. Mutations in the central scaffolding components of NPCs are associated with genetic diseases, but how they manifest only specific tissues remains unclear. This exemplified Nup133-deficient mouse embryonic stem cells, grow normally during pluripotency, differentiate poorly into neurons. Here, using an innovative situ structural biology approach, we show that Nup133 −/− cells have...
The cone-shaped mature HIV-1 capsid is the main orchestrator of early viral replication. After cytosolic entry, it transports replication complex along microtubules toward nucleus. While was initially believed that reverse transcribed genome released from in cytosol, recent observations indicate a high amount protein (CA) remains associated with subviral complexes during import through nuclear pore (NPC). Observation postentry events via microscopic detection CA challenging, since epitope...
The HIV-1 capsid performs essential functions during early viral replication and is an interesting target for novel antivirals. Thus, understanding molecular structural details of function will be important elucidating (and retroviral in general) relevant cells may also aid antiviral development. Here, we show that capsids stay largely intact transport to the nucleus infected T but appear uncoat upon entry into nucleoplasm. These results support hypothesis protect genome from cytoplasmic...
Infection of non-enveloped polyomaviruses depends on an intact microtubular network. Here we focus mouse polyomavirus (MPyV). We show that the dynamics MPyV cytoplasmic transport reflects characteristics motor-driven with bi-directional saltatory movements. In cells treated microtubule-disrupting agents, localization was significantly perturbed, virus retained at cell periphery, mostly within membrane structures resembling multicaveolar complexes, and later times post-infection, only a...
Abstract Human immunodeficiency virus (HIV-1) remains a major health threat. Viral capsid uncoating and nuclear import of the viral genome are critical for productive infection. The size HIV-1 is generally believed to exceed diameter pore complex (NPC), indicating that has occur prior import. Here, we combined correlative light electron microscopy with subtomogram averaging capture structural status reverse transcription-competent complexes in infected T cells. We demonstrate NPC cellulo...
ABSTRACT Murine polyomavirus middle T-antigen (MT) induces tumors by mimicking an activated growth factor receptor. An essential component of this action is a 22-amino-acid hydrophobic region close to the C terminus which locates MT cell membranes. Here, we demonstrate that sequence transmembrane domain (TMD) showing hemagglutinin (HA) tag added exposed on outside cells, with N inside. To determine whether TMD inserted into endoplasmic reticulum (ER) membrane, ER retention signal KDEL...
Minor structural proteins of mouse polyomavirus (MPyV) are essential for virus infection. To study their properties and possible contributions to cell death induction, fusion variants these proteins, created by linking enhanced green fluorescent protein (EGFP) C- or N-termini, were prepared tested in the absence other MPyV gene products, namely tumor antigens major capsid protein, VP1. The minor linked EGFP at C-terminus (VP2-EGFP, VP3-EGFP) found display similar nonfused, wild-type...
Mouse polyomavirus (MPyV) is a member of the Polyomaviridae family, which comprises non-enveloped tumorigenic viruses infecting various vertebrates including humans and causing different pathogenic responses in infected organisms. Despite variations host tropism pathogenicity, structure virions these similar. The capsid, with icosahedral symmetry (ø, 45 nm, T = 7d), composed shell 72 capsomeres structural proteins, arranged around nucleocore containing approximately 5-kbp-long circular dsDNA...
Abstract HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed cDNA molecules their association with cellular proteins using fluorescence correlative-light-and-electron-microscopy (CLEM). specifically detected nuclei, not in cytoplasm. Nuclear initially co-localized fluorescent integrase fusion (IN-FP) CA (capsid) protein, cDNA-punctae separated...
Abstract Chicxulub impact (66 Ma) event resulted in deposition of spheroids and melt glass, followed by diamectite carbonate ejecta represented large polished striated rounded pebbles cobbles, henceforth, called Albion Formation 1 Pook’s Pebbles, name given from the first site identified central Belize, Cayo District. Here we report that magnetic analysis Pebbles samples revealed unique electric discharge signatures. Sectioning at Belize showed different parts had not only contrasting...
The differentiation of colorectal cancer cells is associated with the arrest tumor growth and regression. However, mechanism such cell has not yet been elucidated. Several adenocarcinoma lines, including HT29 which differentiates only upon stimulation a agent, have used for study cells. Since we had previously obtained variable results during analyses these cells, selected several clones this line. In study, four parental H8, G9, G10 A3, were characterized. All them differentiated treatment...
Abstract Gag – the main structural protein of HIV-1 is recruited to plasma membrane for virus assembly by its matrix (MA) domain. subsequently cleaved into component domains, causing maturation repurpose virion cell entry. We determined structure and arrangement MA within immature mature HIV-1, providing a basis understand MA’s role in assembly. Unexpectedly, we found that rearranges during maturation, form new, hexameric lattice which acyl chain phospholipid extends out bind pocket MA. Our...