A5006332749- Cytomegalovirus and herpesvirus research
- Herpesvirus Infections and Treatments
- Toxoplasma gondii Research Studies
- Virus-based gene therapy research
- RNA regulation and disease
- HIV Research and Treatment
- interferon and immune responses
- Viral gastroenteritis research and epidemiology
- Viral Infections and Vectors
- Mosquito-borne diseases and control
- Endoplasmic Reticulum Stress and Disease
- Viral-associated cancers and disorders
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- RNA Interference and Gene Delivery
- Hepatitis B Virus Studies
- Complement system in diseases
- Animal Disease Management and Epidemiology
- Immunotherapy and Immune Responses
- Calcium signaling and nucleotide metabolism
- Viral Infections and Outbreaks Research
- Viral Infectious Diseases and Gene Expression in Insects
- Advanced biosensing and bioanalysis techniques
- Neonatal Health and Biochemistry
- Machine Learning in Bioinformatics
- COVID-19 Impact on Reproduction
Universität Ulm
2007-2024
Institute of Virology of the Slovak Academy of Sciences
2008-2024
University Hospital Ulm
2008-2024
University Medical Center
2015-2024
Cornell University
2004-2009
Friedrich-Loeffler-Institut
2004-2005
New York State College of Veterinary Medicine
2004-2005
Red recombination using PCR-amplified selectable markers is a well-established technique for mutagenesis of large DNA molecules in Escherichia coli. The system has limited efficacy and versatility, however, markerless modifications including point mutations, deletions, particularly insertions longer sequences. Here we describe procedure that combines cleavage with the homing endonuclease I-SceI to allow highly efficient, PCR-based engineering without retention unwanted foreign We applied...
ABSTRACT The innate immune response plays a pivotal role during human cytomegalovirus (HCMV) primary infection. Indeed, HCMV infection of fibroblasts rapidly triggers strong induction type I interferons (IFN-I), accompanied by proinflammatory cytokine release. Here, we show that foreskin (HFFs) infected with mutant TB40/E strain unable to express UL83-encoded pp65 (v65Stop) produce significantly higher IFN-β levels than HFFs the wild-type or revertant (v65Rev), suggesting tegument protein...
ABSTRACT Morphogenesis of human cytomegalovirus (HCMV) is still only partially understood. We have characterized the role HCMV tegument protein pUL71 in viral replication and morphogenesis. By using a rabbit antibody raised against C terminus pUL71, we could detect infected cells, as well virions showing molecular mass approximately 48 kDa. The expression detected early h postinfection, was not blocked by antiviral drug foscarnet, indicating an expression. during virus investigated...
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides target components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish CLR01, a ‘molecular tweezer’ specific lysine arginine residues, inhibits formation of infectivity-enhancing seminal amyloids remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement infection by virion–amyloid complexes...
ABSTRACT A key player in the intrinsic resistance against human cytomegalovirus (HCMV) is interferon-γ-inducible protein 16 (IFI16), which behaves as a viral DNA sensor first hours postinfection and repressor of gene transcription later stages. Previous studies on HCMV replication demonstrated that IFI16 binds to kinase pUL97, undergoes phosphorylation, relocalizes cytoplasm infected cells. In this study, we demonstrate tegument pp65 (pUL83) recruits promoter UL54 downregulates replication,...
The tegument protein pp65 of human cytomegalovirus (HCMV) represents the major component mature virus particles. Nevertheless, deletion has been shown to have no effects on replication and morphogenesis in fibroblasts vitro. We studied HCMV virion composition absence viral growth a stop mutant different cell types, including monocyte-derived macrophages. Two codons at amino acids 11 12 were introduced by bacterial artificial chromosome mutagenesis into endotheliotropic strain TB40/E. Clear...
Electron microscopy (EM) allows visualization of viruses in fixed cells with high resolution. High-pressure freezing for sample fixation combination freeze substitution and embedding resin improves significantly the preservation cellular structures specifically membranes. This advancement better human cytomegalovirus (HCMV) morphogenesis occurring at To obtain comprehensive information on viral phenotypes from ultrastructural images it is important to also quantify morphological phenotypes....
Zika virus (ZIKV) causes severe birth defects and can be transmitted via sexual intercourse. Semen from ZIKV-infected individuals contains high viral loads may therefore serve as an important vector for transmission. Here we analyze the effect of semen on ZIKV infection cells tissues derived anogenital region. replicates in all analyzed cell lines, primary cells, endometrial or vaginal tissues. However, presence semen, by other flaviviruses is potently inhibited. We show that prevents...
Broad-spectrum antivirals are powerful weapons against dangerous viruses where no specific therapy exists, as in the case of ongoing SARS-CoV-2 pandemic. We discovered that a lysine- and arginine-specific supramolecular ligand (CLR01) destroys enveloped viruses, including HIV, Ebola, Zika virus, remodels amyloid fibrils semen promote viral infection. Yet, it is unknown how CLR01 exerts these two distinct therapeutic activities. Here, we delineate novel mechanism antiviral activity by...
The human cytomegalovirus (HCMV) tegument protein pUL71 is required for efficient secondary envelopment and accumulates at the Golgi compartment-derived viral assembly complex (vAC) during infection. Analysis of various C-terminally truncated proteins fused to enhanced green fluorescent (eGFP) identified amino acids 23 34 as important determinants its localization. Sequence analysis mutational verification revealed presence an N-terminal tyrosine-based trafficking motif (YXXΦ) in pUL71. This...
Human cytomegalovirus (HCMV) secondary envelopment requires the viral tegument protein pUL71. The lack of pUL71 results in a complex ultrastructural phenotype with increased numbers capsids undergoing at cytoplasmic virus assembly complex. Here, we report role C terminus envelopment. Mutant viruses expressing C-terminally truncated (TB71del327-361 and TB71del348-351) exhibited an impaired transmission electron microscopy (TEM) studies. Further mutational analyses revealed tetralysine motif...
Abstract Glycoprotein G (gG) of alphaherpesviruses has been described to function as a viral chemokine-binding protein (vCKBP). More recently, mutant viruses devoid gG have shown result in increased virulence, but it remained unclear whether the potential serve vCKBP is responsible for this observation. In study, we used equine herpesvirus type 1 (EHV-1) model study pathophysiological importance activity. First, vitro chemotaxis assays studying migration immune cells, an important...
The product of the human cytomegalovirus (HCMV) UL71 gene is conserved throughout herpesvirus family. During HCMV infection, protein pUL71 required for efficient virion egress and involved in final steps secondary envelopment leading to infectious viral particles. We found strong indications oligomerization under native conditions when recombinant was negatively stained analyzed by electron microscopy. Oligomerization during infection further verified reducing polyacrylamide gel...
The gene M94 of murine cytomegalovirus (MCMV) as well its homologues UL16 in alphaherpesviruses is involved viral morphogenesis. For a better understanding role the life cycle, library random mutants was generated by modified transposon-based linker scanning mutagenesis. A comprehensive set reinserted into MCMV genome and tested for their capacity to complement null mutant. Thereby, 34 loss-of-function were identified, which second screen inhibit virus replication. This analysis identified...
Significance Depending on the host cell type and differentiation status, herpesviruses establish different modes of infection to either maintain or replicate their genomes. How viruses discriminate between individual environments upon is poorly understood. Here we identify a viral sensor mechanism that restricts human cytomegalovirus (HCMV) replication G0/G1 phase division cycle differentiated cells. The based HCMV tegument 150-kDa phosphoprotein which enters as constituent virus particle,...
We report a requirement for the viral protein kinase UL97 in human cytomegalovirus (HCMV) replication that maps to ULb' region of genome. A UL97-null (Δ97) mutant strain TB40/E, which encodes full-length region, exhibited defects, particularly production cell-free virus, were more severe than those seen with Δ97 laboratory AD169, harbors extensive deletions its region. These differences recapitulated additional HCMV strains by treatment inhibitor,...
Human cytomegalovirus (HCMV) encodes an endoplasmic reticulum (ER)-resident glycoprotein, UL148, which activates the unfolded protein response (UPR) but is fully dispensable for viral replication in cultured cells. Hence, its previously ascribed roles immune evasion and modulation of cell tropism are hypothesized to cause ER stress. Here, we show that UL148 necessary sufficient drive formation prominent ER-derived structures on average occupy 5% infected cytoplasm. The sites where coalesces...
Oncolytic viruses (OVs) are promising therapeutics for tumors with a poor prognosis. An OV based on herpes simplex virus type 1 (oHSV-1), talimogene laherparepvec (T-VEC), has been recently approved by the Food and Drug Administration (FDA) European Medicines Agency (EMA) treatment of unresectable melanoma. T-VEC, like most OVs, is administered via intratumoral injection, underlining unresolved problem systemic delivery oncolytic agent metastases deep-seated tumors. To address this drawback,...
ABSTRACT Most equine herpesvirus 1 (EHV-1) strains, including the naturally occurring virulent RacL11 isolate, encode a large glycoprotein, gp2 (250 kDa), which is expressed from gene 71. Besides other alterations in viral genome, avirulent strain KyA harbors an in-frame deletion of 1,242 nucleotides To examine contributions variation to virus growth and virulence, mutant viruses expressing full-length or truncated were generated. Western blot analyses demonstrated expression 250-kDa cells...
Key problems using viral vectors for vaccination and gene therapy are antivector immunity, low transduction efficiencies, acute toxicity, limited capacity to package foreign genetic information. It could be demonstrated that animal human cells were efficiently transduced with equine herpesvirus 1 (EHV-1) reconstituted from DNA maintained manipulated in Escherichia coli. Between 13 23% of primary CD3+, CD4+, CD8+, CD11b+, CD19+ more than 70% CD4+ MT4 or various tumor cell lines (MeWo, Huh7,...