Analysis and nucleotide sequence of an origin of an origin of DNA replication in Acinetobacter calcoaceticus and its use for Escherichia coli shuttle plasmids

Hunger, Michael; Schmucker, Robert; Veerabrahma, Kishan; Hillen, Wolfgang

doi:10.1016/0378-1119(90)90494-c

Cited by 184 publications

(131 citation statements)

References 13 publications

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“…The role of the A. baumannii 19606 csuE-like gene was confirmed further by genetic complementation of the insertion derivative with the parental gene PCR-amplified and cloned in the shuttle vector pWH1266 (Hunger et al, 1990). Electroporation of the recombinant plasmid pMU243 into the A. baumannii 19606 #144 insertion derivative restored its ability to attach and form biofilms in a fashion similar to that displayed by the 19606 parental strain (sample 2 in Fig.…”

Section: Genetic Analysis Of Cell Attachment and Biofilm Formationmentioning

confidence: 80%

Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system

et al. 2003

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Acinetobacter baumannii causes severe infections in compromised patients, survives on abiotic surfaces in hospital environments and colonizes different medical devices. In this study the analysis of the processes involved in surface attachment and biofilm formation by the prototype strain 19606 was initiated. This strain attaches to and forms biofilm structures on plastic and glass surfaces, particularly at the liquid-air interface of cultures incubated stagnantly. The cell aggregates, which contain cell stacks separated by water channels, formed under different culture conditions and were significantly enhanced under iron limitation. Electron and fluorescence microscopy showed that pili and exopolysaccharides are part of the cell aggregates formed by this strain. Electron microscopy of two insertion derivatives deficient in attachment and biofilm formation revealed the disappearance of pili-like structures and DNA sequencing analysis showed that the transposon insertions interrupted genes with the highest similarity to hypothetical genes found in Pseudomonas aeruginosa, Pseudomonas putida and Vibrio parahaemolyticus. Although the products of these genes, which have been named csuC and csuE, have no known functions, they are located within a polycistronic operon that includes four other genes, two of which encode proteins related to chaperones and ushers involved in pili assembly in other bacteria. Introduction of a copy of the csuE parental gene restored the adherence phenotype and the presence of pili on the cell surface of the csuE mutant, but not that of the csuC derivative. These results demonstrate that the expression of a chaperone-usher secretion system, some of whose components appear to be acquired from unrelated sources, is required for pili formation and the concomitant attachment to plastic surfaces and the ensuing formation of biofilms by A. baumannii cells.

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Section: Genetic Analysis Of Cell Attachment and Biofilm Formationmentioning

confidence: 80%

Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system

et al. 2003

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“…Mutagenesis and screening of A. calcoaceticus NCIB8250 as described previously (9) produced strain WH221, which does not grow with phenol as the sole carbon source but which retains the ability to grow at the expense of catechol. A gene library with partially Sau3A-digested chromosomal DNA from the wild type inserted into shuttle plasmid pWH1274 (19) was transformed in WH221 by electroporation and yielded candidates able to grow at the expense of phenol. One recombinant plasmid, called pWH535, contained an insert of 3.0 kb.…”

Section: Resultsmentioning

confidence: 99%

“…The resulting fragments were ligated into XbaI-cleaved, dephosphorylated pQE9 to yield pWH844. XhoI-cleaved, dephosphorylated, and filled-in pWH844 was ligated with the 2.8-kb PvuII-HincII fragment from pWH1266 containing an ori for Acinetobacter (19). The ligation mixture was transformed into A. calcoaceticus ADP1, and transformants were selected on Luria broth (LB) plates with 300 g of ampicillin per ml.…”

Section: Methodsmentioning

confidence: 99%

Expression, inducer spectrum, domain structure, and function of MopR, the regulator of phenol degradation in Acinetobacter calcoaceticus NCIB8250

1997

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“…The complete secA parental allele was PCR amplified using ATCC 19606 T total DNA as a template, Pfu DNA polymerase (Stratagene), and primers 2722 and 2724, both of which included BamHI restriction sites (see Table S1 in the supplemental material). The 3.06-kb amplicon cloned into pCRBlunt II TOPO was subcloned as a BamHI restriction fragment into the cognate site of the A. baumannii-E. coli shuttle vector pWH1266 (32) to generate pMU472 (Table 1; Fig. 1A).…”

Section: Methodsmentioning

confidence: 99%

Role of the Carboxy Terminus of SecA in Iron Acquisition, Protein Translocation, and Virulence of the Bacterial Pathogen Acinetobacter baumannii

et al. 2015

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Acinetobacter baumannii is a Gram-negative opportunistic nosocomial pathogen that causes pneumonia and soft tissue and systemic infections. Screening of a transposon insertion library of A. baumannii ATCC 19606 T resulted in the identification of the 2010 derivative, which, although capable of growing well in iron-rich media, failed to prosper under iron chelation. Genetic, molecular, and functional assays showed that 2010's iron utilization-deficient phenotype is due to an insertion within the 3= end of secA, which results in the production of a C-terminally truncated derivative of SecA. SecA plays a critical role in protein translocation through the SecYEG membrane channel. Accordingly, the secA mutation resulted in undetectable amounts of the ferric acinetobactin outer membrane receptor protein BauA while not affecting the production of other acinetobactin membrane protein transport components, such as BauB and BauE, or the secretion of acinetobactin by 2010 cells cultured in the presence of subinhibitory concentrations of the synthetic iron chelator 2,2=-dipyridyl. Outer membrane proteins involved in nutrient transport, adherence, and biofilm formation were also reduced in 2010. The SecA truncation also increased production of 30 different proteins, including proteins involved in adaptation/tolerance responses. Although some of these protein changes could negatively affect the pathobiology of the 2010 derivative, its virulence defect is mainly due to its inability to acquire iron via the acinetobactin-mediated system. These results together indicate that although the C terminus of the A. baumannii ATCC 19606 T SecA is not essential for viability, it plays a critical role in the production and translocation of different proteins and virulence.

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Analysis and nucleotide sequence of an origin of an origin of DNA replication in Acinetobacter calcoaceticus and its use for Escherichia coli shuttle plasmids

Cited by 184 publications

References 13 publications

Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system

Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system

Expression, inducer spectrum, domain structure, and function of MopR, the regulator of phenol degradation in Acinetobacter calcoaceticus NCIB8250

Role of the Carboxy Terminus of SecA in Iron Acquisition, Protein Translocation, and Virulence of the Bacterial Pathogen Acinetobacter baumannii

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