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151.
Identification of a novel virulence gene, virA, on the large plasmid of Shigella, involved in invasion and intercellular spreading 总被引:4,自引:0,他引:4
152.
The absence of a surface protease, OmpT, determines the intercellular spreading ability of Shigella: the relationship between the ompT and kcpA loci 总被引:12,自引:1,他引:11
N. Nakata T. Tobe I. Fukuda T. Suzuki K. Komatsu M. Yoshikawa C. Sasakawa 《Molecular microbiology》1993,9(3):459-468
A large plasmid-encoded protein, VirG, on the bacterial surface is essential for the spreading of Shigella by eliciting polar deposition of filamentous actin In the cytoplasm of epithelial cells. VirG expression from the large plasmid is diminished greatly when it is introduced into Escherichia coli K-12 from Shigella. In an attempt to identify factors affecting VirG expression, we found that the absence of the ompT gene, encoding outer membrane protease OmpT, restored full production of VirG protein to E. coli K-12. Conversely, upon introduction of the ompT gene of E. coii K-12 into Shigella, spreading ability was completely abolished, probably because of the proteolytic degradation of VirG protein by OmpT. Analysis of the DNA sequence of the ompT region indicated that the absence of the ompT gene occurred in Shigella and enteroinvasive E. coli strains, and that the absent DNA segment corresponded to a remnant lambdoid phage structure found in E. coli K-12, which encompasses a 21 kb DNA segment spanning from argU through to the ompT genes. Since ompT is located near purE in E. coli K-12 and a virulence locus for provoking keratocon-junctivitis in the eyes of guinea-pigs, named kcpA is located near purE in S. fiexnerl, and the two loci are involved in VirG expression, the KcpA~ mutants of S. flexneri 2a constructed were examined for correlation between acquisition of ompT and VirG degradation. Our data suggest that the previous recognition of a kcpA locus in S. flexneri is the result of transfer of the ompr gene from E. coli K-12, giving rise to a KcpA phenotype. These results indicate that the lack of OmpT protease confers upon Shigella the ability to spread into adjacent epithelial cells. 相似文献
153.
154.
155.
Hiroyuki Ohno Tohru Sakai Eishun Tsuchida Kenji Honda Shigeru Sasakawa 《Biochemical and biophysical research communications》1981,102(1):426-431
The membrane fluidity of human erythrocyte ghost was temporarily increased by the addition of polyethylene glycol with molecular weight of 7500. On the other hand, the fluidity of dipalmitoylphosphatidyl choline bilayer liposomes was monotonously decreased by the addition of polyethylene glycol. Fusion of liposomes was inhibited by the interaction with polyethylene glycol. The temporary increase in membrane fluidity of erythrocyte ghosts was considered to be the result of the clustering of membrane-bound proteins which is believed to be one of the most important sequences in cell fusion. 相似文献
156.
Isolation of polyamine transport-deficient mutants of Escherichia coli and cloning of the genes for polyamine transport proteins 总被引:6,自引:0,他引:6
K Kashiwagi N Hosokawa T Furuchi H Kobayashi C Sasakawa M Yoshikawa K Igarashi 《The Journal of biological chemistry》1990,265(34):20893-20897
Escherichia coli KK313, which was deficient in spermidine transport, was isolated by treatment of E. coli MA261 with N-methyl-N'-nitro-N-nitrosoguanidine. E. coli NH1596, which was deficient in spermidine transport and has a 90% decreased putrescine transport activity, was obtained by a second treatment of E. coli KK313 with the same mutagen. Genes for polyamine transport systems were isolated by transforming E. coli NH1596 through DNA fragments from E. coli DR112 using pACYC184 as a vector. One clone for the gene of protein(s) catalyzing both putrescine and spermidine uptake (pPT104) was isolated. Two clones for the genes of protein(s) catalyzing only putrescine uptake (pPT79 and pPT71) were obtained. The genes encoded by pPT104, pPT79, and pPT71 were mapped at 15, 19, and 16 min of E. coli chromosome, respectively. Spermidine uptake by NH1596 carrying pPT104, and by MA261, was not inhibited by putrescine and several polyamine analogues, and the Kt values of these two systems were both approximately 0.1 microM. Putrescine transport by NH1596 carrying pPT104 was inhibited completely by spermidine, N,N-dimethyl-4,4'-bipyridylium (paraquat), and N1-acetyl-spermidine, and the Kt value was 1.4 microM. Putrescine uptake by NH1596 carrying pPT79 or pPT71 was not inhibited by spermidine and several polyamine analogues, and the Kt values were 0.5 and 1.8 microM, respectively. In MA261, the putrescine uptake was inhibited by 25-35% by paraquat and N1-acetyl-polyamines and showed two Kt values, 0.5 and 1.5 microM. Based on these findings, the polyamine transport systems of E. coli are discussed. 相似文献
157.
158.
Transposable elements induce spontaneous mutations, promote genome
rearrangements, regulate gene expression, and participate in the horizontal
spread of genes encoding traits such as antibiotic resistance among
bacterial genera too distantly related to undergo homologous recombination.
Here we review the bacterial transposon Tn5 and focus on those aspects of
its functional organization and transposition which provide insights into
how it and other elements may have arisen, proliferated, and evolved.
相似文献
159.
Identification of citrate utilization transposon Tn3411 from a naturally occurring citrate utilization plasmid. 总被引:13,自引:11,他引:2
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We have isolated a new transposon, Tn3411, encoding citrate-utilizing ability, from a naturally occurring citrate utilization (Cit) plasmid, pOH3001. Citrate transposon Tn3411 was transposed from pOH3001 to lambda b519 b515 cI857 S7 (abbreviated lambda bb) phage, and further from the resulting lambda bb:Tn3411 to a vector plasmid, pBR322, in recA-deficient strains. The Cit+ plasmids (pOH2 and pOH3) constructed by the integration of Tn3411 into pBR322 were examined by restriction endonuclease and heteroduplex analysis. The results obtained were as follows: (i) Tn3411 was 7.4 kilobases long and flanked by small inverted repeats, and it contained one more pair of inverted repeats at the opposite orientation in the internal region, thus making alternate repeats; and (ii) the Cit+ structure gene was located on the fragment (5.5 kilobases) between two SalI cleavage sites on Tn3411. 相似文献
160.
Bacillus subtilis PCI 219 has a single glutamate dehydrogenase (GDH) [EC 1.4.1.3] with dual coenzyme specificity [for NAD(H) and NADP(H)]. The enzyme was purified 800-fold from crude extracts of B. subtilis from the post-exponential phase of growth and showed one significant protein band on gel electrophoresis. This band was determined, by activity staining, to have all the GDH nucleotide specificities. Its molecular weight was estimated to be 250,000+/-20,000 by gel filtration, and 270,000+/-30,000 by zone centrifugation in a sucrose density gradient. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate showed that GDH has a subunit size of about 57,000. The pI of GDH was found to bepH 3.7 by isoelectric focusing. GDH exhibited nonlinear kinetics in the reduction of NAD+, and in the reverse direction, the substrate, NH4+, was strongly inhibitory at high concentrations. Purine nucleotides did not affect the activity. The oxidative demination of glutamate was significantly inhibited by the metabolites oxaloacetate and citrate, which acted as allosteric effectors of this enzyme,inhibiting the reaction in one direction. The pH optimum of each of the activities of GDH and the stability of GDH are also reported. 相似文献