Dokument-ID Dokumenttyp Verfasser/Autoren Herausgeber Haupttitel Abstract Auflage Verlagsort Verlag Erscheinungsjahr Seitenzahl Schriftenreihe Titel Schriftenreihe Bandzahl ISBN Quelle der Hochschulschrift Konferenzname Quelle:Titel Quelle:Jahrgang Quelle:Heftnummer Quelle:Erste Seite Quelle:Letzte Seite URN DOI Abteilungen OPUS4-13405 Wissenschaftlicher Artikel Ondrusch, Nicolai; Kreft, Jürgen Blue and Red Light Modulates SigB-Dependent Gene Transcription, Swimming Motility and Invasiveness in \(Listeria\) \(monocytogenes\) Background: In a number of gram-positive bacteria, including Listeria, the general stress response is regulated by the alternative sigma factor B (SigB). Common stressors which lead to the activation of SigB and the SigB-dependent regulon are high osmolarity, acid and several more. Recently is has been shown that also blue and red light activates SigB in Bacillus subtilis. Methodology/Principal Findings: By qRT-PCR we analyzed the transcriptional response of the pathogen L. monocytogenes to blue and red light in wild type bacteria and in isogenic deletion mutants for the putative blue-light receptor Lmo0799 and the stress sigma factor SigB. It was found that both blue (455 nm) and red (625 nm) light induced the transcription of sigB and SigB-dependent genes, this induction was completely abolished in the SigB mutant. The blue-light effect was largely dependent on Lmo0799, proving that this protein is a genuine blue-light receptor. The deletion of lmo0799 enhanced the red-light effect, the underlying mechanism as well as that of SigB activation by red light remains unknown. Blue light led to an increased transcription of the internalin A/B genes and of bacterial invasiveness for Caco-2 enterocytes. Exposure to blue light also strongly inhibited swimming motility of the bacteria in a Lmo0799- and SigB-dependent manner, red light had no effect there. Conclusions/Significance: Our data established that visible, in particular blue light is an important environmental signal with an impact on gene expression and physiology of the non-phototrophic bacterium L. monocytogenes. In natural environments these effects will result in sometimes random but potentially also cyclic fluctuations of gene activity, depending on the light conditions prevailing in the respective habitat. 2011 e16151 PLoS ONE 6 1 urn:nbn:de:bvb:20-opus-134050 10.1371/journal.pone.0016151 Theodor-Boveri-Institut für Biowissenschaften OPUS4-6384 Wissenschaftlicher Artikel Ondrusch, Nicolai; Kreft, Jürgen Blue and Red Light Modulates SigB-Dependent Gene Transcription, Swimming Motility and Invasiveness in Listeria monocytogenes Background: In a number of gram-positive bacteria, including Listeria, the general stress response is regulated by the alternative sigma factor B (SigB). Common stressors which lead to the activation of SigB and the SigB-dependent regulon are high osmolarity, acid and several more. Recently is has been shown that also blue and red light activates SigB in Bacillus subtilis. Methodology/Principal Findings: By qRT-PCR we analyzed the transcriptional response of the pathogen L. monocytogenes to blue and red light in wild type bacteria and in isogenic deletion mutants for the putative blue-light receptor Lmo0799 and the stress sigma factor SigB. It was found that both blue (455 nm) and red (625 nm) light induced the transcription of sigB and SigB-dependent genes, this induction was completely abolished in the SigB mutant. The blue-light effect was largely dependent on Lmo0799, proving that this protein is a genuine blue-light receptor. The deletion of lmo0799 enhanced the red-light effect, the underlying mechanism as well as that of SigB activation by red light remains unknown. Blue light led to an increased transcription of the internalin A/B genes and of bacterial invasiveness for Caco-2 enterocytes. Exposure to blue light also strongly inhibited swimming motility of the bacteria in a Lmo0799- and SigB-dependent manner, red light had no effect there. Conclusions/Significance: Our data established that visible, in particular blue light is an important environmental signal with an impact on gene expression and physiology of the non-phototrophic bacterium L. monocytogenes. In natural environments these effects will result in sometimes random but potentially also cyclic fluctuations of gene activity, depending on the light conditions prevailing in the respective habitat. 2011 urn:nbn:de:bvb:20-opus-75451 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5095 Wissenschaftlicher Artikel Goebel, Werner; Kreft, Jürgen Accumulation of replicative intermediates and catenated forms of the colicinogenic factor E\(_1\) in E. coli during the replication at elevated temperatures No abstract available 1972 urn:nbn:de:bvb:20-opus-60625 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5094 Wissenschaftlicher Artikel Härtlein, Michael; Schiessl, Sigrid; Wagner, Wilma; Rdest, Ursula; Kreft, Jürgen; Goebel, Werner Transport of hemolysin by Escherichia coli No abstract available 1983 urn:nbn:de:bvb:20-opus-60619 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5049 Wissenschaftlicher Artikel Kreft, Jürgen; Burger, Klaus J.; Goebel, Werner Expression of antibiotic resistance genes from Escherichia coli in Bacillus subtilis Bifunctional recombinant plasmids were constructed, comprised of the E. coli vectors pBR322, pBR325 and pACYC184 and different plasmids from Gram-positive bacteria, e.g. pBSU161-1 of B. subtilis and pUB110 and pC221 of S. aureus. The beta-lactamase (bla) gene and the chloramphenicol acetyltransferase (cat) gene from the E. coli plasmids were not transcribed and therefore not expressed in B. subtilis. However, tetracycline resistance from the E. coli plasmids was expressed in B. subtilis. Transcription of the tetracycline resistance gene(s) started in B. subtilis at or near the original E. coli promoter, the sequence of which is almost identical with the sequence recognized by σ55 of B. subtilis RNA polymerase. 1983 urn:nbn:de:bvb:20-opus-60600 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5084 Wissenschaftlicher Artikel Kreft, Jürgen; Berger, Harald; Härtlein, Michael; Müller, Bodo; Weidinger, Gerhard; Goebel, Werner Cloning and expression in Escherichia coli and Bacillus subtilis of the hemolysin (cereolysin) determinant from Bacillus cereus From a cosmid gene bank of Bacillus cereus GP4 in Escherichia coli we isolated clones which, after several days of incubation, formed hemolysis zones on erythrocyte agar plates. These clones contained recombinant cosmids with B. cereus DNA insertions of varying lengths which shared some common restriction fragments. The smallest insertionwas recloned as aPstl fragment into pJKK3-1, a shuttle vector which repücates in Bacillus subtilis and E. coli. When this recombinant plasmid (pJKK3-1 hly-1) was transformed into E. coli, it caused hemolysis on erythrocyte agar plates, but in liquid assays no extemal or intemal hemolytic activity could be detected with the E. coli transformants. B. subtilis carrying the same plasmid exhibited hemolytic activity at Ievels comparable to those ofthe B. cereus donor strain. The hemolysin produced in B. subtilis seemed to be indistinguishable from cereolysin in its sensitivity to cholesterol, activation by dithiothreitol, and inactivation by antibodies raised against cereolysin. When the recombinant DNA carrying the cereolysin gene was used as a probe in hybridization experiments with chromosomal DNA from a streptolysin 0-producing strain of Streptococcus pyogenes or from üsteriolysin-producing strains of Usteria monoeytogenes, no positive hybridization signals were obtained. These data soggest that the genes for these three SH-activated cytolysins do not have extended sequence homology. 1983 urn:nbn:de:bvb:20-opus-60596 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5071 Wissenschaftlicher Artikel Gilmore, Michael S.; Cruz-Rodz, Armando L.; Leimeister-Wächter, Michaela; Kreft, Jürgen; Goebel, Werner A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage A cloned cytolytic determinant from the genome of Bacillus cereus GP-4 has been characterized at the molecular Ievel. Nucleotide sequence determination revealed the presence of two open reading frames. 8oth open reading frames were found by deletion and complementation analysis to be necessary for expression of the hemolytic phenotype by Bacillus subtilis and Escherichia coli hosts. The 5' open reading frame was found to be nearly identical to a recently reported phospholipase C gene derived from a mutant B. cereus strain which overexpresses the respective protein, and it conferred a lecithinase-positive phenotype to the B. subtilis host. The 3' open reading frame encoded a sphingomyelinase. The two tandemly encoded activities, phospholipase C and sphingomyelinase, constitute a biologically functional cytolytic determinant of B. cereus termed cereolysin AB. 1989 urn:nbn:de:bvb:20-opus-60588 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5070 Wissenschaftlicher Artikel Schülein, Ralf; Kreft, Jürgen; Gonski, Sigrid; Goebel, Werner Preprosubtilisin Carlsberg processing and secretion is blocked after deletion of amino acids 97-101 in the mature part of the enzyme During an investigation into the substrate specificity and processing of subtilisin Carlsberg from Bacillus licheniformis, two major independent findings were made: (i) as has been shown previously, a stretch of five amino acids (residues 97-101 of the mature enzyme) that loops out into the binding cleft is involved in substrate binding by subtilisin Carlsberg. In order to see whether this loop element also determines substrate specificity, the coding region for these five amino acids was deleted from the cloned gene for subtilisin Carlsberg by site-directed mutagenesis. Unexpectedly the resulting mutant preproenzyme (P42c, Mr=42 kDa) was not processed to the mature form (Mr = 30 kDa) and was not released into the medium by a proteasedeficient B. subtilis host strain; rather, it accumulated in the cell membrane. This result demonstrates that the integrity of this loop element, which is very distant from the processing cleavage sites in the preproenzyme, is required for secretion of subtilisin Carlsberg. (ii) In culture supernatants from B. subtilis harbouring the cloned wild-type subtilisin Carlsberg gene the transient appearance (at 0-3 h after onset of stationary phase) of a processing intermediate (P38c, Mr = 38 kDa) oftbis protease could be demonstrated. P38c very probably represents a genuine proform of subtilisin Carlsberg. 1991 urn:nbn:de:bvb:20-opus-60577 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5069 Wissenschaftlicher Artikel Goebel, Werner; Kathariou, S.; Kuhn, M.; Sokolovic, Z.; Kreft, Jürgen; Köhler, S.; Funke, D.; Chakraborty, T.; Leimeister-Wächter, M. Hemolysin from Listeria-biochemistry, genetics and function in pathogenesis No abstract available 1988 urn:nbn:de:bvb:20-opus-60563 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5068 Wissenschaftlicher Artikel Goebel, Werner; Chakraborty, T.; Kreft, Jürgen Bacterial hemolysins as virulence factors No abstract available 1988 urn:nbn:de:bvb:20-opus-60553 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5067 Wissenschaftlicher Artikel Kreft, Jürgen; Funke, Dorothee; Haas, Albert; Lottspeich, Friedrich; Goebel, Werner Production, purification and characterization of hemolysins from Listeria ivanovii and Listeria monocytogenes Sv4b. In culture supematants of both Listeria ivanovii and Listeria monocytogenes Sv4b, for the first time a hemolysin of molecular weight 58 kDa was identified, which had all the characteristics of an SH-activated cytolysin, and which was therefore identified as Iisteriolysin 0 (LLO). In the case of L. ivanovii a second major supematant protein of molecular weight 24 kDa co-purified with LLO. However, the function of this protein has to be determined. In culture supematants of L. ivanovii a sphingomyelinase and a Iecithinase activity could be detected, both enzymatic activities together contributing to the pronounced hemolysis caused by L. ivanovii. The N-tenninal amino acid sequences of LLO and the 24 kDa from L. ivanovii are shown. 1989 urn:nbn:de:bvb:20-opus-60545 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5047 Wissenschaftlicher Artikel Haas, Albert; Brehm, Klaus; Kreft, Jürgen; Goebel, Werner Cloning, characterization, and expression in Escherichia coli of a gene encoding Listeria seeligeri catalase, a bacterial enzyme highly homologous to mammalian catalases A gene coding for catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase; EC 1.11.1.6) of the grain-positive bacterium Listeria seeligeri was cloned from a plasmid library of EcoRI-digested chromosomal DNA, with Escherichia coli DHSa as a host. The recombinant catalase was expressed in E. coli to an enzymatic activity approximately SO times that of the combined E. coli catalases. The nucleutide sequence was determined, and the deduced amino acid sequence revealed 43.2% amino acid sequence identity between bovine liver catalase and L. seeligeri catalase. Most of the amino acid residues which are involved in catalytic activity, the formation of the active center accession channel, and heme binding in bovine liver catalase were also present in L. seeligeri catalase at the corresponding positions. The recombinant protein contained 488 amino acid residues and had a calculated molecular weight of 55,869. The predicted isoelectric point was 5.0. Enzymatic and genetic analyses showed that there is most probably a single catalase of this type in L. seeligeri. A perfect 21-bp inverted repeat, which was highly homologous to previously reported binding sequences of the Fur (ferric uptake regulon) protein of E. coli, was detected next to the putative promoter region of tbe L. seeligeri catalase gene. 1991 urn:nbn:de:bvb:20-opus-60536 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5046 Wissenschaftlicher Artikel Haas, Albert; Dumbsky, Martina; Kreft, Jürgen Listeriolysin genes: complete sequence of ilo from Listeria ivanovii and of lso from Listeria seeligeri The completc DNA scqucnccs coding for thc thiol-activated cytolysins from Listeria ivanovii, ivanolysin 0 (ILO) and for sccligerolysin 0 (LSO) from Listeria seeligeri have been dctermined. Thc deduced amino acid scquences revealed that: (i) the primary translation products comprise 528 (ILO) and 530 (LSO) amino acids. respectively. (ii) ILO contains two cysteines. LSO has a substitution in the conserved cysteine motif. 1992 urn:nbn:de:bvb:20-opus-60529 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5045 Wissenschaftlicher Artikel Brehm, Klaus; Haas, Albert; Goebel, Werner; Kreft, Jürgen A gene encoding a superoxide dismutase of the facultative intracellular bacterium Listeria monocytogenes A gene (Imsod) encoding superoxide dismutase (SOD; EC 1.15.1.1) of the facultative intracellular pathogen, Listeria monocytogenes, was cloned by functional complementation of an SOD-deficient Escherichia coli mutant. The nucleotide sequence was determined and the deduced amino acid (aa) sequence (202 aa) showed close similarity to manganese-containing SOD's from other organisms. Subunits of the recombinant L. monocytogenes SOD (re-SOD) and of both E. coli SODs formed enzymatically active hybrid enzymes in vivo. DNA/DNA-hybridization experiments showed that this type of recombinant re-sod gene is conserved within the genus Listeria. 1992 urn:nbn:de:bvb:20-opus-60515 Theodor-Boveri-Institut für Biowissenschaften OPUS4-5044 Wissenschaftlicher Artikel Lampidis, Robert; Gross, Roy; Sokolovic, Zeljka; Goebel, Werner; Kreft, Jürgen The virulence regulator protein of Listeria ivanovii is highly homologous to PrfA from Listeria monocytogenes and both belong to the Crp-Fnr family of transcription regulators No abstract available 1994 urn:nbn:de:bvb:20-opus-60503 Theodor-Boveri-Institut für Biowissenschaften OPUS4-4089 Wissenschaftlicher Artikel Kreft, Jürgen; Haas, Albert; Goebel, Werner Isolation and characterization of genes coding for proteins involved in the cytolysis by Listeria ivanovii We established a library of chromosomal DNA of Listeria ivanovii in the pTZ19R plasmid system, using Escherichia coli DH5alpha as the host. One recombinant clone reacted strongly with a polyclonal antiserum raised against the listeriolysin 0 and a second exoprotein (24kDa) of L. ivanovii, which is most probably also involved in cytolytic processes. The recombinant E. coli clone may contain part of the listeriolysin 0 gene of L. ivanovii. 1989 urn:nbn:de:bvb:20-opus-46991 Theodor-Boveri-Institut für Biowissenschaften OPUS4-4088 Wissenschaftlicher Artikel Kreft, Jürgen; Funke, D.; Schlesinger, R.; Lottspeich, F.; Goebel, Werner Purification and characterization of cytolysins from Listeria monocytogenes serovar 4b and Listeria ivanovii Several exoproteins from Listeria monocytogenes serovar 4b (NCTC 10527) and Listeria ivanovii (ATCC) 19119, SLCC 2379), respectively, have been purified to homogeneity by thiol-disulfide exchange chromatography and gel filtration. Both strains produce a haemolytic/cytolytic protein of Mr 58 kDa, which has all the properties of a SH-activated cytolysin, the prototype of which is streptolysin 0 (SLO), and this protein has therefore heen termed Iisteriolysin 0 (LLO). In addition a protein of Mr 24 kDa from culture supernatants of L. ivanovii co-purified withLLO. The N-terminal aminoacid sequences of both proteins from L. ivanovii have been determined. By mutagenesis with transposons of Gram-positive origin (Tn916 and TnI545), which have been introduced via conjugation into L. ivanovii, several phenotypic mutants (altered haemolysis on sheep blood agar or lecithinase-negative) were obtained. Results on the properties of these muntants will he presented. 1989 urn:nbn:de:bvb:20-opus-47036 Theodor-Boveri-Institut für Biowissenschaften OPUS4-4087 Wissenschaftlicher Artikel Kreft, Jürgen; Hughes, Colin Cloning vectors derived from plasmids and phage of Bacillus No abstract available 1982 urn:nbn:de:bvb:20-opus-47014 Theodor-Boveri-Institut für Biowissenschaften OPUS4-4086 Wissenschaftlicher Artikel Kreft, Jürgen; Bernhard, K.; Goebel, Werner Recombinant plasmids capable to replication in B. subtilis and E. coli The plasmid pBC16 (4.25 kbases), ongtnally isolated from Bacillus cereus, determines tetracycline resistance and can be transformed into competent cells of B. subtilis. A miniplasmid of pBCl6 (pBCI6-1), 2,7 kb) which has lost an EcoRI fragment of pBCI6 retains the replication functions and the tetracycline resistance. This plasmid which carries only one EcoRI site has been joined in vitro to pBS], a cryptic plasmid previously isolated from B. subtilis and shown to carry also a single EcoRI site (Bernhard et aI., 1978). The recombinant plasmid is unstable and dissociates into the plasmid pBSl61 (8.2 kb) and the smaller plasmid pBS162 (2. I kb). Plasmid pBS161 retains the tetracycline resistance. It possesses a single EcoRI site and 6 HindlII sites. The largest HindIII fragment of pBS161 carries the tetracycline resistance gene and the replication function. After circularization in vitro of this fragment a new plasmid, pBS161-l is generated, which can be used as a HindlII and EcoRI cloning vector in Bacillus suhtilis. Hybrid plasmids consisting of the E. coli plasmids pBR322, p WL 7 or pACl84 and different HindlII fragments of pBSI61 were constructed in vitro. Hybrids containing together with the E. coli plasmid the largest HindlII fragment of pBS161 can replicate in E. coli and B. sublilis. In E. coli only the replicon of the E. coli plasmid part is functioning whereas in B. suhtilis replication of the hybrid plasmid is under the control of the Bacillus replicon. The tetracycline resistance of the B. subtilis plasmid is expressed in E. coli, but several antibiotic resistances of the E. coli plasmids (ampicillin, kanamycin and chloramphenicol) are not expressed in B. suhtilis. The hybrid plasmids seem to be more unstable in B. subtilis than in E. coli. 1978 urn:nbn:de:bvb:20-opus-47000 Theodor-Boveri-Institut für Biowissenschaften OPUS4-4084 Wissenschaftlicher Artikel Kreft, Jürgen Reovirus-specific messenger ribonucleoprotein particles from Hela cells When reovirus-infected Hela cells are incubated at 43°C virus-specific messenger RNA is released ~rom the polysomes. It accumulates free in the cytoplasm as messenger ribonucleoprotem partIcles (mRNPs). The:e part~cles have a sedimentati~n rate of about 50S and a buoyant densIty m CsCI of 1.42 g/cm . ReovIrus mRNPs contam, beSIdes all three size classes of reovirus messenger RNA, the same spectrum of proteins found in the polysomal mRNPs from uninfected cells, plus t~o addi~ional pr?teins with molecular masses of 7000~ d and 110000 d, respectively. Electron mIcroscoPIc exammatlOn of the reovIrus mRNP fractIOn reveals specific Y-shaped structures wIth a total mean length ofO.5Ilm. 1980 urn:nbn:de:bvb:20-opus-47028 Theodor-Boveri-Institut für Biowissenschaften