A5077903795- Bacterial Genetics and Biotechnology
- Bacterial Infections and Vaccines
- Bacteriophages and microbial interactions
- DNA Repair Mechanisms
- Pneumonia and Respiratory Infections
- Streptococcal Infections and Treatments
- Monoclonal and Polyclonal Antibodies Research
- Biochemical and Structural Characterization
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Antibiotic Resistance in Bacteria
- Legume Nitrogen Fixing Symbiosis
- Plant tissue culture and regeneration
- Escherichia coli research studies
- Transgenic Plants and Applications
- Helicobacter pylori-related gastroenterology studies
- Complement system in diseases
- CRISPR and Genetic Engineering
- Advanced Biosensing Techniques and Applications
- Clostridium difficile and Clostridium perfringens research
- Glycosylation and Glycoproteins Research
- Genomics and Phylogenetic Studies
- Antimicrobial Resistance in Staphylococcus
- Cassava research and cyanide
- vaccines and immunoinformatics approaches
Toscana Life Sciences
2009-2023
Novartis (Italy)
2006-2014
Novartis (Germany)
2008-2013
Novartis (Switzerland)
2006-2013
CS Diagnostics
2013
Leiden University
1998-2001
Max Planck Institute for Molecular Genetics
1986-2000
Centro Nacional de Biotecnología
2000
Consejo Superior de Investigaciones Científicas
2000
Max Planck Society
1992-1996
The IncP alpha promiscuous plasmid (R18, R68, RK2, RP1 and RP4) comprises 60,099 bp of nucleotide sequence, encoding at least 74 genes. About 40 kb the genome, designated core including all essential replication transfer functions, can be aligned with equivalent sequences in beta R751. compiled sequence revealed several previously unidentified reading frames that are potential plasmids carry genetic information very efficiently: coding genes closely packed but rarely overlap, occupy almost...
Analysis of publicly available genomes Streptococcus pneumoniae has led to the identification a new genomic element containing genes typical gram-positive pilus islets (PIs). Here, we demonstrate that this region, herein referred as PI-2 (consisting pitA, sipA, pitB, srtG1, and srtG2) codes for second functional in pneumococcus. Polymerization requires backbone protein PitB well sortase SrtG1 signal peptidase-like SipA. Presence correlates with genotype defined by multilocus sequence typing...
Transfer genes of the IncP plasmid RP4 are grouped in two separate regions, designated Tra1 and Tra2. Tra2 gene products proposed to be mainly responsible for formation mating pairs conjugating cells. To provide information relevant understanding function products, nucleotide sequence entire region is presented here. Twelve open reading frames were identified core region, being essential intraspecific Escherichia coli matings. Predicted sizes 11 12 polypeptides could verified by expression...
As an early stage of plant transformation by Agrobacterium tumefaciens, the Ti plasmid is nicked at border sequences that delimit T-DNA. Cleavage results in covalent attachment VirD2 to 5' terminal strand a process resembling initiation DNA transfer occurs donor cell during bacterial conjugation. We demonstrate this cleavage can be reproduced vitro: protein, border-cleaving enzyme, was overproduced and purified. assays were performed with single-stranded oligodeoxyribonucleotides...
We have constructed a RP4 KorB overproducing strain and purified the protein to near homogeneity. is DNA binding recognizing defined paiindromic 13-bp sequences (TTTAGCSGCTAAA). Inverted sequence repetitions of this type, designated OB, are present on 12 times. O B -sequences localized In replication maintenance regions as well in Tra1 Tra2 essential for conjugative transfer. All sites found Tra by computer search act targets specific protein. KorB-DNA complexes were detected fragment...
During initiation of conjugative transfer DNA containing the origin (oriT) promiscuous plasmid RP4, proteins TraI, TraJ, and TraH interact assemble a specialized nucleoprotein complex (the relaxosome) at oriT. The structure can be visualized on electron micrographs. Site- strand-specific nicking in vitro is dependent TraI TraJ Mg2+ ions. Substrate specificity directed exclusively towards cognate origin: RP4-specified protein cannot recognize closely related oriT R751. After nicking, remains...
Streptococcus pneumoniae is a major public health threat worldwide. The recent discovery that this pathogen possesses pili led us to investigate their protective abilities in mouse model of intraperitoneal infection. Both active and passive immunization with recombinant pilus subunits afforded protection against lethal challenge the S. serotype 4 strain TIGR4.
To characterize protein-DNA interactions involved in the initiation of conjugative transfer replication, we isolated and sequenced origins (oriT) promiscuous IncP plasmids RP4 R751. The central initiating event at origin a plasmid is cleavage unique site (nic) strand to be transferred recipient cell. This process can triggered after assembly "relaxosomes" (plasmid DNA-protein relaxation complexes), requiring plasmid-encoded gene products. We analyzed nicking reaction for demonstrated that...
Journal Article Common sequence motifs in DNA relaxases and nick regions from a variety of transfer systems Get access Werner Pansegrau, Pansegrau Max-Planck-Institut för Molekulare Genetik, Abteilung Schuster, Ihnestrasse73, D-1 000 Berlin 33, FRG Search for other works by this author on: Oxford Academic PubMed Google Scholar Erich Lanka * *To whom correspondence should be addressed Nucleic Acids Research, Volume 19, Issue 12, 25 June 1991, Page 3455, https://doi.org/10.1093/nar/19.12.3455...
Conjugative DNA transfer of the self-transmissible broad-host-range plasmid RP4 is initiated by strand- and site-specific cleavage at nick site (nic) origin (oriT). Cleavage results in covalent attachment plasmid-encoded relaxase (TraI) to 5'-terminal 2'-deoxycytidine residue nic. We demonstrate that Tyr22 center catalytic TraI, mediating via formation a phosphodiester between 5' phosphoryl aromatic hydroxyl group. The specificity seen with form I oriT was verified short...
RP4 TrbB, an essential component of the conjugative transfer apparatus broad-host-range plasmid RP4, is a member PulE protein superfamily involved in multicomponent machineries transporting macromolecules across bacterial envelope. PulE-like proteins share several well conserved motifs, most notable nucleoside triphosphate binding motif (P-loop). Helicobacter pylori HP0525 also belongs to and encoded by pathogenicity island cag, inflammatory response infected gastric epithelial cells...
The IncP antibiotic-resistance plasmids transfer to a broad range of bacterial species. RK2 origin DNA (oriT) consists 250-base-pair segment including the single-stranded cleavage site (nic) needed generate strand believed be transferred. Deletion derivatives and bank hydroxylamine-generated oriT mutants were screened for loss transferability. regions flanking both sides nic are required optimal clone. Of chemically induced mutants, critical base-pair changes that dramatically reduced...
The TraI protein of plasmid RP4 (IncP alpha) catalyzes a site- and strand-specific cleaving-joining reaction on form I or single-stranded DNA. Thus, is one the key components involved in initiation termination horizontal DNA transfer by bacterial conjugation. Amino acid sequence comparison revealed three motifs conserved relaxases from different origins. Site-directed mutagenesis traI structural gene application purified mutant proteins for vitro assays served to evaluate functional...
Formation of relaxosomes is the first step in initiation transfer DNA replication during bacterial conjugation. This nucleoprotein complex contains all components capable introducing a site- and strand-specific nick at cognate origin (oriT) on supercoiled plasmid DNA, thus providing substrate for generation strand to be transferred. Characterization terminal nucleotides oriT site revealed that relaxation occurs by hydrolysis single phosphodiester bond between 2'-deoxyguanosyl 2'-deoxycytidyl...
Abstract Systematic characterisation of the reactivity lysine moieties in CRM 197 towards N ‐hydroxysuccinimide linkers bearing alkynes or azides is described. This involves two‐step conjugation various glycans to by click chemistry a well‐defined manner. By semiquantitative LC‐MS/MS analysis proteolytic digests conjugates formed, residues protein was mapped and ranked. Computational solvent accessibility each residue (based on crystal structure) established correlation between surface...
4CMenB is the first broad coverage vaccine for prevention of invasive meningococcal disease caused by serogroup B strains. To gain a comprehensive picture antibody response induced upon vaccination and to obtain relevant translational information directly from human studies, we have isolated panel monoclonal antibodies adult vaccinees. Based on Ig-gene sequence variable region, 37 antigen-specific were identified produced as recombinant Fab fragments, subset also full length IgG1...
T-DNA processing during agroinfection of plants is initiated by site- and strand-specific incision at the border sequences Ti plasmid. Two proteins are required for this reaction: VirD2 (49.6 kDa), catalyzing a site-specific cleaving-joining reaction on single-stranded DNA in vitro (Pansegrau, W., Schoumacher, F., Hohn, B., Lanka, E.(1993) Proc. Natl. Acad. Sci. U. S. A. 90, 11538-11542), VirD1 (16.1 an accessory protein VirD2-mediated specific cleavage double-stranded DNA. Following...
ABSTRACT Type IV secretion systems direct transport of protein or nucleoprotein complexes across the cell envelopes prokaryotic donor and eukaryotic recipient cells. The process is mediated by a membrane-spanning multiprotein assembly. Potential NTPases belonging to VirB11 family are an essential part complex. Three representatives these originating from conjugative transfer regions plasmids RP4 (TrbB) R388 (TrwD) cag pathogenicity island Helicobacter pylori (HP0525) were overproduced...
Two essential transfer genes of the conjugative plasmid RP4 were altered by site-directed mutagenesis: traG primase operon and traI relaxase operon. To evaluate effects on phenotype point mutations, we have reconstituted system fusion regions Tra1 Tra2 to small multicopy replicon ColD. Deletions in or served determine Tra mutant plasmids trans complementation. motifs TraG which are highly conserved among TraG-like proteins several other DNA systems found be for function. One resembles that a...
The nucleotide sequence of the relaxase operon and leader which are part Tral region promiscuous plasmid RP4 was determined. These two polycistronic operons transcribed divergently from an intergenic about 360 bp containing transfer origin six close-packed genes. A seventh gene completely overlaps another one in a different reading frame. Conjugative DNA proceeds unidirectio nally oriT with heading to be transferred. tral includes within its 3′ terminal promoter controlling 7.2-kb primase...
Staphylococcal protein A (SpA) is a multifunctional, highly conserved virulence factor of