@article{GilmoreCruzRodzLeimeisterWaechteretal.1989,
author = {Gilmore, Michael S. and Cruz-Rodz, Armando L. and Leimeister-W{\"a}chter, Michaela and Kreft, J{\"u}rgen and Goebel, Werner},
title = {A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60588},
year = {1989},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{BrehmHaasGoebeletal.1992,
author = {Brehm, Klaus and Haas, Albert and Goebel, Werner and Kreft, J{\"u}rgen},
title = {A gene encoding a superoxide dismutase of the facultative intracellular bacterium Listeria monocytogenes},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60515},
year = {1992},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{GoebelKreft1972,
author = {Goebel, Werner and Kreft, J{\"u}rgen},
title = {Accumulation of replicative intermediates and catenated forms of the colicinogenic factor E\(_1\) in E. coli during the replication at elevated temperatures},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60625},
year = {1972},
abstract = {No abstract available},
subject = {Biologie},
language = {en}
}
@article{GoebelChakrabortyKreft1988,
author = {Goebel, Werner and Chakraborty, T. and Kreft, J{\"u}rgen},
title = {Bacterial hemolysins as virulence factors},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60553},
year = {1988},
abstract = {No abstract available},
subject = {Biologie},
language = {en}
}
@article{GoetzEylertEisenreichetal.2010,
author = {Goetz, Andreas and Eylert, Eva and Eisenreich, Wolfgang and Goebel, Werner},
title = {Carbon Metabolism of Enterobacterial Human Pathogens Growing in Epithelial Colorectal Adenocarcinoma (Caco-2) Cells},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68555},
year = {2010},
abstract = {Analysis of the genome sequences of the major human bacterial pathogens has provided a large amount of information concerning their metabolic potential. However, our knowledge of the actual metabolic pathways and metabolite fluxes occurring in these pathogens under infection conditions is still limited. In this study, we analysed the intracellular carbon metabolism of enteroinvasive Escherichia coli (EIEC HN280 and EIEC 4608-58) and Salmonella enterica Serovar Typhimurium (Stm 14028) replicating in epithelial colorectal adenocarcinoma cells (Caco-2). To this aim, we supplied [U-13C6]glucose to Caco-2 cells infected with the bacterial strains or mutants thereof impaired in the uptake of glucose, mannose and/or glucose 6-phosphate. The 13C-isotopologue patterns of protein-derived amino acids from the bacteria and the host cells were then determined by mass spectrometry. The data showed that EIEC HN280 growing in the cytosol of the host cells, as well as Stm 14028 replicating in the Salmonella-containing vacuole (SCV) utilised glucose, but not glucose 6-phosphate, other phosphorylated carbohydrates, gluconate or fatty acids as major carbon substrates. EIEC 4608-58 used C3-compound(s) in addition to glucose as carbon source. The labelling patterns reflected strain-dependent carbon flux via glycolysis and/or the Entner-Doudoroff pathway, the pentose phosphate pathway, the TCA cycle and anapleurotic reactions between PEP and oxaloacetate. Mutants of all three strains impaired in the uptake of glucose switched to C3-substrate(s) accompanied by an increased uptake of amino acids (and possibly also other anabolic monomers) from the host cell. Surprisingly, the metabolism of the host cells, as judged by the efficiency of 13C-incorporation into host cell amino acids, was not significantly affected by the infection with either of these intracellular pathogens.},
subject = {Metabolismus},
language = {en}
}
@article{KreftBergerHaertleinetal.1983,
author = {Kreft, J{\"u}rgen and Berger, Harald and H{\"a}rtlein, Michael and M{\"u}ller, Bodo and Weidinger, Gerhard and Goebel, Werner},
title = {Cloning and expression in Escherichia coli and Bacillus subtilis of the hemolysin (cereolysin) determinant from Bacillus cereus},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60596},
year = {1983},
abstract = {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 rep{\"u}cates 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 {\"u}steriolysin-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.},
subject = {Biologie},
language = {en}
}
@article{BergerHackerJuarezetal.1982,
author = {Berger, Harald and Hacker, J{\"o}rg and Juarez, Antonio and Hughes, Colin and Goebel, Werner},
title = {Cloning of the chromosomal determinants encoding hemolysin production and mannose-resistant hemagglutination in Escherichia coli},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40255},
year = {1982},
abstract = {We have cloned the chromosomal hemolysin determinants from Escherichia coli strains belonging to the four O-serotypes 04, 06, 018, and 075, The hemolysin-producing clones were isolated from gene banks of these strains which were constructed by inserting partial Sau3A fragments of chromosomal DNA into the cosmid pJC74. The hemolytic cosmid clones were relatively stable. The inserts were further sub cloned either as Sail fragments in pACYC184 or as BamHI-SaLI fragments in a recombinant plasmid (pANN202) containing cistron C (hlye) of the plasmid-encoded hemolysin determinant. Detailed restriction maps of each of these determinants were constructed, and it was found that, despite sharing overall homology, the determinants exhibited minor specific differences in their structure, These appeared to be restricted to cistron A (hlyA), which is the structural gene for hemolysin. In the gene banks of two of these hemolytic strains, we could also identify clones which carried the genetic determinants for the mannose-resistant hemagglutination antigens Vb and VIc. Both of these fimbrial antigens were expressed in the E. coli K-12 clones to an extent similar to that observed in the wild-type strains. These recombinant cosmids were rather unstable, and, in the absence of selection, segregated at a high frequency.},
language = {en}
}
@article{HaasBrehmKreftetal.1991,
author = {Haas, Albert and Brehm, Klaus and Kreft, J{\"u}rgen and Goebel, Werner},
title = {Cloning, characterization, and expression in Escherichia coli of a gene encoding Listeria seeligeri catalase, a bacterial enzyme highly homologous to mammalian catalases},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60536},
year = {1991},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{KreftGoebel1974,
author = {Kreft, J{\"u}rgen and Goebel, Werner},
title = {Complex Co1E1 DNA in Escherichia coli and Proteus mirabilis},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47044},
year = {1974},
abstract = {Incubation of the colicinogenic Escherichia coli strain JC 411 (ColE1) at elevated temperatures (47-49°) leads to the accumulation of catenated molecules and replicative intermediates of this plasmid. Mature supercoiled OolE1 DNA molecules synthesized under these conditions have an increased number of tertiary turns as shown by electron microscopy. The monomeric tightly supercoiled molecules possess a slightly slower sedimentation rate and a higher binding capacity for ethidium bromide than supercoiJed monomers synthesized at lower temperatures. Recombination deficient mutants of E. coli recA, recB and recC, which carry the ColE1 plasmid, form about the same amount of catenated molecules at the elevated temperature as a rec+ strain. In addition, we have observed by electron microscopy a small percentage (.--.5\% of the circular DNA molecules) of minicircular DNA molecules in all preparations of JC 411 (CoIE1). They are homogenous in size, with a molecular weight of 1.4 X 106 daltons. Addition of chloramphenicol to a culture of Proteus mirabilis (ColE1) leads to an increased amount of higher multiple circular oligomers and to a stimulated accumulation of catenated OolE1 DNA molecules of varying sizes. ColE1 DNA synthesis is more thermosensitive than chromosomal DNA replication in P. mirabili8. Plasmid replication stops completely at temperatures above 43°C.},
language = {en}
}
@article{KreftBurgerGoebel1983,
author = {Kreft, J{\"u}rgen and Burger, Klaus J. and Goebel, Werner},
title = {Expression of antibiotic resistance genes from Escherichia coli in Bacillus subtilis},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60600},
year = {1983},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{HughesMuellerHackeretal.1982,
author = {Hughes, Colin and M{\"u}ller, Dorothee and Hacker, J{\"o}rg and Goebel, Werner},
title = {Genetics and pathogenic role of Escherichia coli hemolysin},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40082},
year = {1982},
abstract = {While clear evidence exists for the direct involvement of cytolysins in the pathogenesis of Gram-positive bacteria, the significance of Gram-negative haemolysins remains unclear. This paper presents briefly data indicating a role for haemolysin production in infections caused by Escherichia coli and also experiments which have allowed an analysis of the molecular basis of the haemolysis among pathogenic and non-pathogenic strains of this species.},
language = {en}
}
@article{HackerOttBlumetal.1992,
author = {Hacker, J{\"o}rg and Ott, Manfred and Blum, Gabriele and Marre, Reinhard and Heesemann, J{\"u}rgen and Tsch{\"a}pe, Helmut and Goebel, Werner},
title = {Genetics of Escherichia coli uropathogenicity: Analysis of the O6:K15:H31 isolate 536},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-71578},
year = {1992},
abstract = {E. coli strain 536 (06: K15: H31) isolated from a case of acute pyelonephritis, expresses S-fimbrial adhesins, P-related fimbriae, common type I fimbriae, and hemolysins. The respective chromosomally encoded determinants were cloned by constructing a genomic library of this strain. Furthermore, the strain produces the iron uptake substance, enterocheline, damages HeLa cells, and behaves in a serum-resistant mode. Genetic analysis of spontaneously arising non-hemolytic variants revealed that some of the virulence genes were physically linked to large unstable DNA regions, termed "pathogenicity islands", which were mapped in the respective positions on the E. coli K-12linkage map. By comparing the wild type strain and mutants in in vitro and in vivo assays, virulence features have been evaluated. In addition, a regulatory cross talk between adhesin determinants was found for the wild-type isolate. This particular mode of virulence regulation is missing in the mutant strain.},
subject = {Escherichia coli},
language = {en}
}
@article{GoebelKathariouKuhnetal.1988,
author = {Goebel, Werner and Kathariou, S. and Kuhn, M. and Sokolovic, Z. and Kreft, J{\"u}rgen and K{\"o}hler, S. and Funke, D. and Chakraborty, T. and Leimeister-W{\"a}chter, M.},
title = {Hemolysin from Listeria-biochemistry, genetics and function in pathogenesis},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60563},
year = {1988},
abstract = {No abstract available},
subject = {Biologie},
language = {en}
}
@article{EisenreichRudelHeesemannetal.2019,
author = {Eisenreich, Wolfgang and Rudel, Thomas and Heesemann, J{\"u}rgen and Goebel, Werner},
title = {How viral and intracellular bacterial pathogens reprogram the metabolism of host cells to allow their intracellular replication},
series = {Frontiers in Cellular and Infection Microbiology},
volume = {9},
journal = {Frontiers in Cellular and Infection Microbiology},
issn = {2235-2988},
doi = {10.3389/fcimb.2019.00042},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-197188},
year = {2019},
abstract = {Viruses and intracellular bacterial pathogens (IBPs) have in common the need of suitable host cells for efficient replication and proliferation during infection. In human infections, the cell types which both groups of pathogens are using as hosts are indeed quite similar and include phagocytic immune cells, especially monocytes/macrophages (MOs/MPs) and dendritic cells (DCs), as well as nonprofessional phagocytes, like epithelial cells, fibroblasts and endothelial cells. These terminally differentiated cells are normally in a metabolically quiescent state when they are encountered by these pathogens during infection. This metabolic state of the host cells does not meet the extensive need for nutrients required for efficient intracellular replication of viruses and especially IBPs which, in contrast to the viral pathogens, have to perform their own specific intracellular metabolism to survive and efficiently replicate in their host cell niches. For this goal, viruses and IBPs have to reprogram the host cell metabolism in a pathogen-specific manner to increase the supply of nutrients, energy, and metabolites which have to be provided to the pathogen to allow its replication. In viral infections, this appears to be often achieved by the interaction of specific viral factors with central metabolic regulators, including oncogenes and tumor suppressors, or by the introduction of virus-specific oncogenes. Less is so far known on the mechanisms leading to metabolic reprogramming of the host cell by IBPs. However, the still scant data suggest that similar mechanisms may also determine the reprogramming of the host cell metabolism in IBP infections. In this review, we summarize and compare the present knowledge on this important, yet still poorly understood aspect of pathogenesis of human viral and especially IBP infections.},
language = {en}
}
@article{KreftHaasGoebel1989,
author = {Kreft, J{\"u}rgen and Haas, Albert and Goebel, Werner},
title = {Isolation and characterization of genes coding for proteins involved in the cytolysis by Listeria ivanovii},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-46991},
year = {1989},
abstract = {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.},
language = {en}
}
@article{ChakrabortyKathariouHackeretal.1987,
author = {Chakraborty, Trinad and Kathariou, Sophia and Hacker, J{\"o}rg and Hof, Herbert and Huhle, Burkhard and Wagner, Wilma and Kuhn, Michael and Goebel, Werner},
title = {Molecular analysis of bacterial cytolysins},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40328},
year = {1987},
abstract = {Results of molecular and pathogenic studies of three different bacterial hemolysins (cytolysins) are presented. These exoproteins derive from the two gram-negative bacteria Escherichia coli and Aeromonas hydrophila and from the gram-positive pathogen Listeria monocytogenes. The hemolysin of E. coli is determined by an 8-kilobase (kb) region that includes four clustered genes (hlyC, hlyA, hlyB, and hlyD). This hemolysin determinant is part either of large transmissible plasmids or of the chromosome. The genes located chromosomally are found predominantly in E. coli strains that can cause pyelonephritis and/or other extraintestinal infections. A detailed analysis of the chromosomal hly determinants of one nephropathogenic E. coli strain revealed the existence of specific, large chromosomal insertions 75 kb and lOO kb in size that carry the hly genes but that also influence the expression of other virulence properties, i.e., adhesion and serum resistance. The direct involvement of E. coli hemolysin in virulence could be demonstrated in several model systems. The genetic determinants for hemolysin (cytolysin) formation in , A. hydrophila (aerolysin) and L. monocytogenes (listeriolysin) are less complex. Both cytolysins seem to be encoded by single genes, although two loci (aerB and aerC) that affect the expression and activity of aerolysin have been identified distal and proximal to the structural gene for aerolysin (aerA). Cytolysin-negative mutants of both bacteria were obtained by site-specific deletion and/or transposon mutagenesis. These mutants show a drastic reduction in the virulence of the respective bacteria.},
language = {en}
}
@article{KnappHackerThenetal.1984,
author = {Knapp, Stefan and Hacker, J{\"o}rg and Then, Irene and M{\"u}ller, Dorothee and Goebel, Werner},
title = {Multiple copies of hemolysin genes and associated sequences in the chromosome of uropathogenic Escherichia coli strains},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-40278},
year = {1984},
abstract = {The 06 serogroup Escherichia coli strain 536 carries two hemolysin (hly) determinants integrated into the chromosome. The two hly determinants are not completely identical, either functionally or structurally, as demonstrated by spontaneous deletion mutants carrying only one of them and by cloning each of the two determinants separately into cosmid vectors. Each hly determinant is independently deleted at a frequency of 10-4 , leading to variants which exhibit similar levels of internal hemolysin but different amounts of secreted hemolysin. The two hly determinants were also identified in the 04 E. coli strain 519. The three E. coli strains 251, 764, and 768, which belong to the serogroup 018, and the 04 strain 367 harbor a single chromosomal hly determinant, as demonstrated by hybridization with hly-gene-specific probes. However, a hybridization probe derived from a sequence adjacent to the hlyC-proximal end of the plasmid pHlyl52-encoded hly determinant hybridizes with several additional chromosomal bands in hemolytic 018 and 06 E. coli strains and even in E. coli K-12. The size ofthe probe causing the multiple hybridization suggests a 1,500- to 1,800-base pair sequence directly flanking hlyC. Spontaneous hemolysin-negative mutants were isolated from strains 764 and 768, which had lost the entire hly determinant but retained all copies of the hlyC-associated sequence. This sequence is not identical to a previously identified (J. Hacker, S. Knapp, and W. Goebel, J. Bacteriol. 154:1145-1154, 1983) somewhat smaller (about 850 base pairs) sequence flanking the other (hlyBb-proximal) end of the plasmid pHlyl52-encoded hly determinant which, as shown here, exists also in multiple copies in these hemolytic E. coli strains and in at least two copies in E. coli K-12. In contrast to the plasmid-encoded hly determinant which is directly flanked at both ends by these two diJJerent sequences, the chromosomal hly determinants are not immediately flanked by such sequences.},
language = {en}
}
@article{EisenreichRudelHeesemannetal.2021,
author = {Eisenreich, Wolfgang and Rudel, Thomas and Heesemann, J{\"u}rgen and Goebel, Werner},
title = {Persistence of Intracellular Bacterial Pathogens—With a Focus on the Metabolic Perspective},
series = {Frontiers in Cellular and Infection Microbiology},
volume = {10},
journal = {Frontiers in Cellular and Infection Microbiology},
issn = {2235-2988},
doi = {10.3389/fcimb.2020.615450},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-222348},
year = {2021},
abstract = {Persistence has evolved as a potent survival strategy to overcome adverse environmental conditions. This capability is common to almost all bacteria, including all human bacterial pathogens and likely connected to chronic infections caused by some of these pathogens. Although the majority of a bacterial cell population will be killed by the particular stressors, like antibiotics, oxygen and nitrogen radicals, nutrient starvation and others, a varying subpopulation (termed persisters) will withstand the stress situation and will be able to revive once the stress is removed. Several factors and pathways have been identified in the past that apparently favor the formation of persistence, such as various toxin/antitoxin modules or stringent response together with the alarmone (p)ppGpp. However, persistence can occur stochastically in few cells even of stress-free bacterial populations. Growth of these cells could then be induced by the stress conditions. In this review, we focus on the persister formation of human intracellular bacterial pathogens, some of which belong to the most successful persister producers but lack some or even all of the assumed persistence-triggering factors and pathways. We propose a mechanism for the persister formation of these bacterial pathogens which is based on their specific intracellular bipartite metabolism. We postulate that this mode of metabolism ultimately leads, under certain starvation conditions, to the stalling of DNA replication initiation which may be causative for the persister state.},
language = {en}
}
@article{SchueleinKreftGonskietal.1991,
author = {Sch{\"u}lein, Ralf and Kreft, J{\"u}rgen and Gonski, Sigrid and Goebel, Werner},
title = {Preprosubtilisin Carlsberg processing and secretion is blocked after deletion of amino acids 97-101 in the mature part of the enzyme},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60577},
year = {1991},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{KreftFunkeHaasetal.1989,
author = {Kreft, J{\"u}rgen and Funke, Dorothee and Haas, Albert and Lottspeich, Friedrich and Goebel, Werner},
title = {Production, purification and characterization of hemolysins from Listeria ivanovii and Listeria monocytogenes Sv4b.},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60545},
year = {1989},
abstract = {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.},
subject = {Biologie},
language = {en}
}
@article{KreftFunkeSchlesingeretal.1989,
author = {Kreft, J{\"u}rgen and Funke, D. and Schlesinger, R. and Lottspeich, F. and Goebel, Werner},
title = {Purification and characterization of cytolysins from Listeria monocytogenes serovar 4b and Listeria ivanovii},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47036},
year = {1989},
abstract = {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.},
language = {en}
}
@article{KreftBernhardGoebel1978,
author = {Kreft, J{\"u}rgen and Bernhard, K. and Goebel, Werner},
title = {Recombinant plasmids capable to replication in B. subtilis and E. coli},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-47000},
year = {1978},
abstract = {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.},
language = {en}
}
@article{HeisigFrentzenBergmannetal.2011,
author = {Heisig, Martin and Frentzen, Alexa and Bergmann, Birgit and Gentschev, Katharina Ivaylo and Hotz, Christian and Schoen, Christoph and Stritzker, Jochen and Fensterle, Joachim and Rapp, Ulf R. and Goebel, Werner},
title = {Specific antibody-receptor interactions trigger InlAB-independent uptake of Listeria monocytogenes into tumor cell lines},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68705},
year = {2011},
abstract = {Background: Specific cell targeting is an important, yet unsolved problem in bacteria-based therapeutic applications, like tumor or gene therapy. Here, we describe the construction of a novel, internalin A and B (InlAB)-deficient Listeria monocytogenes strain (Lm-spa+), which expresses protein A of Staphylococcus aureus (SPA) and anchors SPA in the correct orientation on the bacterial cell surface. Results: This listerial strain efficiently binds antibodies allowing specific interaction of the bacterium with the target recognized by the antibody. Binding of Trastuzumab (Herceptin®) or Cetuximab (Erbitux®) to Lm-spa+, two clinically approved monoclonal antibodies directed against HER2/neu and EGFR/HER1, respectively, triggers InlABindependent internalization into non-phagocytic cancer cell lines overexpressing the respective receptors. Internalization, subsequent escape into the host cell cytosol and intracellular replication of these bacteria are as efficient as of the corresponding InlAB-positive, SPA-negative parental strain. This specific antibody/receptormediated internalization of Lm-spa+ is shown in the murine 4T1 tumor cell line, the isogenic 4T1-HER2 cell line as well as the human cancer cell lines SK-BR-3 and SK-OV-3. Importantly, this targeting approach is applicable in a xenograft mouse tumor model after crosslinking the antibody to SPA on the listerial cell surface. Conclusions: Binding of receptor-specific antibodies to SPA-expressing L. monocytogenes may represent a promising approach to target L. monocytogenes to host cells expressing specific receptors triggering internalization.},
subject = {Listeria monocytogenes},
language = {en}
}
@article{LampidisGrossSokolovicetal.1994,
author = {Lampidis, Robert and Gross, Roy and Sokolovic, Zeljka and Goebel, Werner and Kreft, J{\"u}rgen},
title = {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},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60503},
year = {1994},
abstract = {No abstract available},
subject = {Biologie},
language = {en}
}
@article{HaertleinSchiesslWagneretal.1983,
author = {H{\"a}rtlein, Michael and Schiessl, Sigrid and Wagner, Wilma and Rdest, Ursula and Kreft, J{\"u}rgen and Goebel, Werner},
title = {Transport of hemolysin by Escherichia coli},
url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-60619},
year = {1983},
abstract = {No abstract available},
subject = {Biologie},
language = {en}
}