Organisation, Potential Regulatory Elements and Evolution of trp Genes in Acinetobacter

Haspel, G.; Veerabrahma, Kishan; Hillen, Wolfgang

doi:10.1007/978-1-4899-3553-3_16

Cited by 6 publications

(6 citation statements)

References 27 publications

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“…4A), which is relatively far apart compared with the optimal spacing in E. coli (17 bp [24]). Interestingly, a 19-bp spacing has also been found for the trpFB operon of A. calcoaceticus BD413 (23). The Ϫ35 region of the putative lipB promoter is also identical to the equivalent region upstream of the A. calcoaceticus trpE gene.…”

Section: Vol 177 1995 a Lipase Chaperone In A Calcoaceticus 3301mentioning

confidence: 75%

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Characterization of lipase-deficient mutants of Acinetobacter calcoaceticus BD413: identification of a periplasmic lipase chaperone essential for the production of extracellular lipase

Kok¹,

Thor²,

Nugteren-Roodzant³

et al. 1995

J Bacteriol

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At the onset of the stationary phase in rich media, the gram-negative soil bacterium Acinetobacter calcoaceticus BD413 produces several lipolytic enzymes (40), which have partly overlapping substrate specificities. BD413 estA, encoding one of the cell-bound esterases (40), and lipA, encoding the extracellular lipase (43), have been cloned and characterized. The regulation of their expression is currently being investigated. In addition, we also aim at defining posttranscriptional factors involved in regulation of enzyme production. The latter aspect is especially important for the extracellular lipase of A. calcoaceticus BD413, since several components involved in export of the protein, including concurrent protein processing events, may have significant control over lipase production.The extracellular lipase of A. calcoaceticus BD413 presumably is exported from the cell via a two-step translocation process (43): across the cytoplasmic membrane, this translocation will be mediated by the Sec system (59, 66), whereas translocation through the outer membrane proceeds via a translocation complex similar to the Xcp system in Pseudomonas spp. (69) and the Pul system in Klebsiella spp. (59). These assumptions are based upon the identification of an N-terminal export signal peptide, indicative of the involvement of a Seclike system, and upon the high degree of similarity of A. calcoaceticus LipA to Pseudomonas lipases that are secreted via a two-step mechanism. Moreover, we have demonstrated that a periplasmic processing enzyme, a protein disulfide oxidoreductase, is required for high-level production of lipase in A. calcoaceticus BD413 (43).In Pseudomonas spp., production of lipases that are similar to Acinetobacter LipA requires the presence of a lipase-specific helper protein, which is N terminally anchored in the cytoplasmic membrane, with its C-terminal domain in the periplasm. Activity of this type of protein is essential for the second lipase translocation step, across the outer membrane (14, 34). In the periplasm, such helper proteins have been shown to mediate the formation of secondary structure elements in the periplasmic form of the lipase, suggesting a chaperone-like function of the helper protein. Unlike the situation in pseudomonads, as yet no essential lipase helper protein has been identified in A. calcoaceticus. In Pseudomonas species, these proteins are invariably encoded downstream of the structural lipase gene, and this is not the case in A. calcoaceticus BD413 (43).Previously, we have reported on the identification of two

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Section: Vol 177 1995 a Lipase Chaperone In A Calcoaceticus 3301mentioning

confidence: 75%

“…The Ϫ35 region of the putative lipB promoter is also identical to the equivalent region upstream of the A. calcoaceticus trpE gene. However, this region is separated from its consensus Ϫ10 region (TATAAT) by only 16 nucleotides (23).…”

Section: Vol 177 1995 a Lipase Chaperone In A Calcoaceticus 3301mentioning

confidence: 99%

Characterization of lipase-deficient mutants of Acinetobacter calcoaceticus BD413: identification of a periplasmic lipase chaperone essential for the production of extracellular lipase

Kok¹,

Thor²,

Nugteren-Roodzant³

et al. 1995

J Bacteriol

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“…The clustering of these catabolic genes in Acinetobacter sp. ADPl is contrasted by the genomic separation of the trp genes needed for tryptophan biosynthesis (Haspel et al, 1991). In E. coli, there are five tryptophan genes, trpEDCBA, which are contiguous in one operon, with the trpD and trpC genes encoding bifunctional enzymes.…”

Section: Genome Size and Genetic Organizationmentioning

confidence: 99%

“…In P. aeruginosa, there are at least four linkage groups, trpE, trpGDC, trpF and trpBA, which are found in three well-separated chromosomal locations (Haspel et al, 1991;Holloway et al, 1990;Schmidt et al, 1996).…”

Section: Genome Size and Genetic Organizationmentioning

confidence: 99%

Directed introduction of DNA cleavage sites to produce a high-resolution genetic and physical map of the Acinetobacter sp. strain ADP1 (BD413UE) chromosome

1997

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The natural transformability of the soil bacterium Acinetobacter sp. ADPl (BD413UE), formerly classified as A. calcoaceticus, has facilitated previous physiological and biochemical investigations. In the present studies, the natural transformation system was exploited to generate a physical and genetic map of this strain's 378021 91 kbp circular chromosome. Previously isolated Acinetobacter genes were modified in vitro to incorporate a recognition sequence for the restriction endonuclease Notl. Following transformation of the wild-type strain by the modified DNA, homologous recombination placed each engineered Not1 cleavage site at the chromosomal location of the corresponding gene. This allowed precise gene localization and orientation of more than 40 genes relative to a physical map which was constructed with transverse alternating field electrophoresis (TAFE) and Southern hybridization methods. The positions of Notl, Ascl and I-Ceul recognition sites were determined, and the latter enzyme identified the presence of seven ribosomal RNA operons. Multiple chromosomal copies of insertion sequence IS1236 were indicated by hybridization. Several of these copies were concentrated in one region of the chromosome in which a spontaneous deletion of approximately 100 kbp occurred. Moreover, contrary to previous reports, ColEl-based plasmids appeared to replicate autonomously in Acinetobacter sp. ADPI.

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“…For example, intracellular gene conversion can give rise to strains that exhibit substantial genetic instability (Doten et ai, 1987a), and it should be possible to determine if such instability is maintained in organisms lacking a functional recA gene. In addition, rec/4-deficient strains should allow the construction of stable merodiploids and thus facilitate analysis of cytoplasmic interactions between regulatory gene products and their chromosomal targets (Cohn and Crawford, 1976;Haspei et al. 1991;Neidie et ai.…”

Section: Introduction Of the Reca100tn5 Mutation By Transformationmentioning

confidence: 99%

Properties of Acinetobacter calcoaceticus recA and its contribution to intracellular gene conversion

1994

Molecular Microbiology

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The Acinetobacter calcoaceticus pcaJ and catJ genes, nearly identical in DNA sequence, differ in transcriptional control and are separated by more than 20 kb of chromosomal DNA. The pcaJ3125 mutation is repaired frequently in organisms containing the wild-type catJ gene. This high-frequency repair is eliminated in strains lacking the catJ gene, which suggests that recombination between the homologous catJ and pcaJ genes contributes to the high-frequency repair of the pcaJ3125 mutation. We report here that the high-frequency repair also requires a functional recA gene. The A. calcoaceticus recA gene was cloned in Escherichia coli by complementation of a recA mutation in the host strain. The nucleotide sequence of a 1506 bp DNA fragment containing A. calcoaceticus recA was determined. The amino acid sequences of RecA from E. coli and A. calcoaceticus shared 71% identity. The DNA sequences differed in that a consensus binding site for binding of LexA repressor, represented upstream from recA in E. coli, is not evident in the corresponding region of the A. calcoaceticus DNA sequence. A Tn5 insertion was introduced into the A. calcoaceticus recA gene. Selection for Tn5-encoded kanamycin resistance allowed the inactivated recA gene to be recombined by natural transformation into the A. calcoaceticus chromosome. Strains that had acquired the mutant gene were sensitive to both MMS and u.v. light, were deficient in natural transformation, and failed to carry out catJ-dependent high-frequency repair of the pcaJ3125 mutation.

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Organisation, Potential Regulatory Elements and Evolution of trp Genes in Acinetobacter

Cited by 6 publications

References 27 publications

Characterization of lipase-deficient mutants of Acinetobacter calcoaceticus BD413: identification of a periplasmic lipase chaperone essential for the production of extracellular lipase

Characterization of lipase-deficient mutants of Acinetobacter calcoaceticus BD413: identification of a periplasmic lipase chaperone essential for the production of extracellular lipase

Directed introduction of DNA cleavage sites to produce a high-resolution genetic and physical map of the Acinetobacter sp. strain ADP1 (BD413UE) chromosome

Properties of Acinetobacter calcoaceticus recA and its contribution to intracellular gene conversion

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