Transductional analysis of chloramphenicol biosynthesis genes in Streptomyces venezuelae

Vats, Siddharth; Stuttard, C.; Vining, L. C.

doi:10.1128/jb.169.8.3809-3813.1987

Cited by 34 publications

(19 citation statements)

References 5 publications

(6 reference statements)

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“…3). This location for tyr-2 is consistent with its earlier map assignment between hisA6 and thrC1 (Chaliciopoulos, 1985;Vats et al, 1987), and is comparable with that of tyrA,B in S. coelicolor A3(2) (Hopwood & Kieser, 1990). In crosses with VS161, which carries the tyr-2 marker, no segregation was observed between tyr-2+ and tsr of AP41; other markers in each cross segregated independently.…”

Section: Chromosomal Location Of Trpegsupporting

confidence: 75%

Location of the genes for anthranilate synthase in Streptomyces venezuelae ISP5230: genetic mapping after integration of the cloned genes

Paradkar

Stuttard²,

Vining³

1993

Journal of General Microbiology

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The anthranilate synthase (trpEG) genes in Streptomyces venezuefae ISP5230 were located by allowing a segregationally unstable plasmid carrying cloned S. venezuefae trpEG DNA and a thiostrepton resistance (tsr) marker to integrate into the chromosome. The integrated tsr was mapped by conjugation and transduction to a location close to tyr-2, between arg-6 and trpAI3. A genomic DNA fragment containing trpC from S. venezuefae ISP5230 was cloned by complementation of a trpC mutation in Streptomyces fividans. Evidence from restriction enzyme analysis of the cloned DNA fragments, from Southern hybridization using the cloned trp DNA as probes, and from cotransduction frequencies, placed trpEG at a distance of 12-45 kb from the trpCBA cluster. The overall arrangement of tryptophan biosynthesis genes in the S. venezuefae chromosome differs from that in other bacteria examined so far.

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Section: Chromosomal Location Of Trpegsupporting

confidence: 75%

Location of the genes for anthranilate synthase in Streptomyces venezuelae ISP5230: genetic mapping after integration of the cloned genes

Paradkar

Stuttard²,

Vining³

1993

Journal of General Microbiology

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“…Consistent with these expectations, Brown et al (1996) demonstrated that Cm production in Streptomyces venezuelae ISP5230 was severely impaired when an indigenous gene that complemented a Streptomyces lividans pab mutant was replaced with a disrupted allele. The locus has not yet been genetically linked to the mapped Cm biosynthesis gene cluster in S. venezuelae ISP5230 (Doull et al, 1986 ;Vats et al, 1987), but the DNA that complemented the S. lividans pab lesion has been shown by cloning and sequence analysis to be a gene consisting of fused pabA and pabB domains. In their deduced amino acid sequences, these domains closely resemble the products of discrete pabA and pabB genes in a variety of bacteria (Brown et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

p-Aminobenzoic acid and chloramphenicol biosynthesis in Streptomyces venezuelae: gene sets for a key enzyme, 4-amino-4-deoxychorismate synthase The GenBank accession number for the sequence reported in this paper is AF189258.

Chang

Sun

et al. 2001

Microbiology

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Amplification of sequences from Streptomyces venezuelae ISP5230 genomic DNA using PCR with primers based on conserved prokaryotic pabB sequences gave two main products. One matched pabAB, a locus previously identified in S. venezuelae. The second closely resembled the conserved pabB sequence consensus and hybridized with a 3 8 kb NcoI fragment of S. venezuelae ISP5230 genomic DNA. Cloning and sequence analysis of the 3 8 kb fragment detected three ORFs, and their deduced amino acid sequences were used in BLAST searches of the GenBank database. The ORF1 product was similar to PabB in other bacteria and to the PabB domain encoded by S. venezuelae pabAB. The ORF2 product resembled PabA of other bacteria. ORF3 was incomplete ; its deduced partial amino acid sequence placed it in the MocR group of GntR-type transcriptional regulators. Introducing vectors containing the 3 8 kb NcoI fragment of S. venezuelae DNA into pabA and pabB mutants of Escherichia coli, or into the Streptomyces lividans pab mutant JG10, enhanced sulfanilamide resistance in the host strains. The increased resistance was attributed to expression of the pair of discrete translationally coupled p-aminobenzoic acid biosynthesis genes (designated pabB/pabA) cloned in the 3 8 kb fragment. These represent a second set of genes encoding 4-amino-4-deoxychorismate synthase in S. venezuelae ISP5230. In contrast to the fused pabAB set previously isolated from this species, they do not participate in chloramphenicol biosynthesis, but like pabAB they can be disrupted without affecting growth on minimal medium. The gene disruption results suggest that S. venezuelae may have a third set of genes encoding PABA synthase.

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“…Those for chloramphenicol are flanked by the auxotrophic markers pdx-4 and cys-28 (Doull et al, 1986 ;Vats et al, 1987 ;Stuttard, 1988). Our initial interest in the cys-28 marker led us to explore the molecular biology and biochemistry of cysteine biosynthesis in Str.…”

Section: Filamentous Bacteria Of the Genusmentioning

confidence: 99%

“…venezuelae ISP5230 (Vats et al, 1987) was examined for growth requirements imposed by the unidentified cys-28 mutation. When colonies on MM agar were supplemented with substances potentially involved in cysteine biosynthesis, positive responses were obtained with cysteine, methionine, homocysteine and cystathionine (Table 2).…”

Section: Test For Complementation Of the Cys-28 Mutation With Orf1mentioning

confidence: 99%

Biosynthesis of sulfur-containing amino acids in Streptomyces venezuelae ISP5230: roles for cystathionine β-synthase and transsulfuration b bThe GenBank accession number for the sequence reported in this paper is AF319543.

Chang

Vining

2002

Microbiology

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A 0 5 kb fragment of Streptomyces venezuelae ISP5230 genomic DNA was amplified by PCR using primers based on consensus sequences of cysteine synthase isozyme A from bacteria. The deduced amino acid sequence of the PCR product resembled not only cysteine synthase sequences from prokaryotes and eukaryotes but also eukaryotic cystathionine β-synthase sequences.Probing an Str. venezuelae genomic library with the PCR product located a hybridizing colony from which pJV207 was isolated. Sequencing and analysis of the Str. venezuelae DNA insert in pJV207 detected two ORFs. The deduced amino acid sequence of ORF1 matched both cysteine synthase and cystathionine β-synthase sequences in GenBank, but its size favoured assignment as a cystathionine β-synthase. ORF2 in the pJV207 insert was unrelated in function to ORF1 ; in its sequence the deduced product resembled acetyl-CoA transferases, but disruption of the ORF did not cause a detectable phenotypic change. Disruption of ORF1 failed to elicit cysteine auxotrophy in wild-type Str. venezuelae, but in the cys-28 auxotroph VS263 it prevented restoration of prototrophy with homocysteine or methionine supplements. The change in phenotype implicated loss of the transsulfuration activity that in the wild-type converts these supplements to cysteine. This study concludes that disruption of ORF1 inactivates a cbs gene, the product of which participates in cysteine synthesis by transsulfuration. Enzyme assays of Str. venezuelae mycelial extracts confirmed the formation of cysteine by thiolation of Oacetylserine, providing the first unambiguous detection of this activity in a streptomycete. Enzyme assays also detected cystathionine γ-synthase, cystathionine β-lyase and cystathionine γ-lyase activity in the extracts and showed that the substrate for cystathionine γ-synthase was O-succinylhomoserine. Based on assay results, the cys-28 mutation in Str. venezuelae VS263 does not inactivate the cysteine synthase gene but impairs expression in cultures grown in minimal medium.

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Transductional analysis of chloramphenicol biosynthesis genes in Streptomyces venezuelae

Cited by 34 publications

References 5 publications

Location of the genes for anthranilate synthase in Streptomyces venezuelae ISP5230: genetic mapping after integration of the cloned genes

Location of the genes for anthranilate synthase in Streptomyces venezuelae ISP5230: genetic mapping after integration of the cloned genes

p-Aminobenzoic acid and chloramphenicol biosynthesis in Streptomyces venezuelae: gene sets for a key enzyme, 4-amino-4-deoxychorismate synthase The GenBank accession number for the sequence reported in this paper is AF189258.

Biosynthesis of sulfur-containing amino acids in Streptomyces venezuelae ISP5230: roles for cystathionine β-synthase and transsulfuration b bThe GenBank accession number for the sequence reported in this paper is AF319543.

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