Use of polymerase chain reaction to identify a leucyl tRNA in Streptomyces coelicolor

Trepanier, Nicole K; N'Guyen, G. Duchateau; Leedell, Paul J.; Leskiw, Brenda K.

doi:10.1016/s0378-1119(97)00077-2

Cited by 15 publications

(27 citation statements)

References 18 publications

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“…3. Similar to analysis of the bldA tRNA from S. coelicolor (Leskiw et al, 1993 ;Trepanier, 1999 ;Trepanier et al, 1997), the bldA tRNA showed a temporal pattern of accumulation in wild-type S. clavuligerus, with higher levels of the tRNA being seen in older, differentiating cultures. This was in contrast to the pattern of accumulation of the processed form of an abundant leucyl-tRNA (encoded by leuU), which was present at relatively constant levels throughout growth.…”

Section: Antibiotics and The Blda Trna Both Show Growthphase Dependensupporting

confidence: 64%

“…RNA was extracted from surfacegrown or liquid cultures as described elsewhere (Leskiw et al, 1993). Northern blot analysis of bldA and leuU transcripts was performed as previously described (Trepanier et al, 1997) [the probes for bldA, leuU and the 5S rRNA control were the oligonucleotides BKL5, BKL42 and BKL53 (Table 2), respectively, except that the hybridization temperature for the bldA probe was 37 mC]. Northern blot analysis of ccaR was performed according to Williams & Mason (1985) as previously described (Leskiw et al, 1993).…”

Section: Methodsmentioning

confidence: 99%

“…Since the processed, mature form of the bldA tRNA showed growth-phase dependent accumulation in S. coelicolor (Leskiw et al, 1993 ;Trepanier et al, 1997), it was of interest to determine if the bldA tRNA would show the same pattern of accumulation in S. clavuligerus. RNA was isolated in duplicate at various time points between 24 and 72 h post-inoculation from liquid cultures, and between 30 and 168 h post-inoculation from surface-grown cultures of S. clavuligerus NRRL 3585 and S. clavuligerus ∆bldA4-1.…”

Section: Antibiotics and The Blda Trna Both Show Growthphase Dependenmentioning

confidence: 99%

“…Although the S. coelicolor bldA tRNA is transcribed from a vegetative promoter and expressed throughout growth, it appears to be regulated at the post-transcriptional level. In contrast to a tRNA for the abundant leucine codon CUC, the mature, 5h-processed bldA tRNA shows growth-phase dependent accumulation, being expressed at low levels until the onset of secondary metabolism and morphological differentiation and reaching maximal levels of accumulation late in growth (Leskiw et al, 1993 ;Trepanier et al, 1997). Consistent with the low level of the tRNA Leu UUA in vegetative cultures, a TTA-containing reporter gene, ampC, showed delayed production of β-lactamase in S. lividans (Leskiw et al, 1991b), and a subtilisin-inhibitor-encoding ssi gene, that had been engineered to contain two TTA codons, showed reduced expression, particularly during early growth, in S. lividans (Ueda et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

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The positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus is mistranslated in a bldA mutant The GenBank accession number for the sequence reported in this paper is AF436078.

et al. 2002

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In Streptomyces coelicolor bldA encodes the principal leucyl tRNA for translation of UUA codons and controls pigmented antibiotic production by the presence of TTA codons in the genes encoding the pathway-specific activators of actinorhodin and undecylprodigiosin biosynthesis. In Streptomyces clavuligerus the gene encoding the pathway-specific activator of both cephamycin C and clavulanic acid production, ccaR, also contains a TTA codon and was expected to exhibit bldA-dependence. A cloned S. clavuligerus DNA fragment containing a sequence showing 91 % identity to the S. coelicolor bldA-encoded tRNA was able to restore antibiotic production and sporulation to bldA mutants of S. coelicolor and the closely related Streptomyces lividans. A null mutation of the bldA gene in S. clavuligerus resulted in the expected sporulation defective phenotype, but unexpectedly had no effect on antibiotic production. Transcript analysis showed no difference in the levels of ccaR transcripts in the wild-type and bldA mutant strains, ruling out any effect of elevated levels of the ccaR mRNA. Furthermore, when compared to the wildtype strain, the bldA mutant showed no differences in the levels of CcaR, suggesting that the single TTA codon in ccaR is mistranslated efficiently. The role of codon context in bldA dependence is discussed.

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Section: Antibiotics and The Blda Trna Both Show Growthphase Dependensupporting

confidence: 64%

Section: Methodsmentioning

confidence: 99%

Section: Antibiotics and The Blda Trna Both Show Growthphase Dependenmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus is mistranslated in a bldA mutant The GenBank accession number for the sequence reported in this paper is AF436078.

et al. 2002

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“…Although they do not impede growth, bldA mutations of S. coelicolor do have phenotypic consequences: the colonies fail to form aerial mycelium on most laboratory media, and they also fail to make several different antibiotics. These processes are typically stationary-phase associated, and indeed the bldA tRNA shows increased abundance in stationary phase, whereas other tRNAs are more abundant during rapid growth (266,437). From such observations, the idea grew that bldA has a regulatory role in relation to the biology of the stationary phase (reviewed in references 66 and 264).…”

Section: P450 Cytochromes (Cypsmentioning

confidence: 99%

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Ventura

Canchaya

Tauch

et al. 2007

Microbiol Mol Biol Rev

946

638

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SUMMARY Actinobacteria constitute one of the largest phyla among Bacteria and represent gram-positive bacteria with a high G+C content in their DNA. This bacterial group includes microorganisms exhibiting a wide spectrum of morphologies, from coccoid to fragmenting hyphal forms, as well as possessing highly variable physiological and metabolic properties. Furthermore, Actinobacteria members have adopted different lifestyles, and can be pathogens (e.g., Corynebacterium, Mycobacterium, Nocardia, Tropheryma, and Propionibacterium), soil inhabitants (Streptomyces), plant commensals (Leifsonia), or gastrointestinal commensals (Bifidobacterium). The divergence of Actinobacteria from other bacteria is ancient, making it impossible to identify the phylogenetically closest bacterial group to Actinobacteria. Genome sequence analysis has revolutionized every aspect of bacterial biology by enhancing the understanding of the genetics, physiology, and evolutionary development of bacteria. Various actinobacterial genomes have been sequenced, revealing a wide genomic heterogeneity probably as a reflection of their biodiversity. This review provides an account of the recent explosion of actinobacterial genomics data and an attempt to place this in a biological and evolutionary context.

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Evolutionary flux of potentially bldA-dependent Streptomyces genes containing the rare leucine codon TTA

Chandra

Chater

2008

Antonie van Leeuwenhoek

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Previous studies have shown that one of the six leucine codons, UUA, is rare in Streptomyces, and that, while the gene for the UUA-specific tRNA, bldA, can generally be inactivated in diverse streptomycetes without impairing vegetative growth, bldA mutants are typically defective in reproductive aerial growth and in antibiotic production. Here, four complete genome sequences and 143 gene clusters for antibiotic biosynthesis from diverse streptomycetes were analysed in order to evaluate the evolution and function of genes whose possession of TTA codons makes them dependent on bldA. It was deduced that the last common ancestor of the four sequenced genomes, possibly 220 million years ago, already possessed the bldA system, together with perhaps 200 TTA-containing target genes. Some 33 of these genes are retained by the modern descendants, though only three of them retain a TTA in all occurrences. Nearly all of these 33, as well as many of the TTA-containing genes with orthologues in two or three of the four genomes, have the same location on the chromosomes as in their common ancestor. However, the majority of TTA-containing genes (61% overall in the four genomes) are species-specific, and were probably acquired by comparatively recent horizontal gene transfer. Most of these genes are of unknown function, and it is likely that many of them confer specialised ecological benefits. On the other hand, one class of species-specific, functionally recognisable, horizontally acquired genes--the gene clusters for antibiotic production--very often contain TTA codons; and nearly half of them have TTA codons in their pathway-specific regulatory genes.

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Use of polymerase chain reaction to identify a leucyl tRNA in Streptomyces coelicolor

Cited by 15 publications

References 18 publications

The positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus is mistranslated in a bldA mutant The GenBank accession number for the sequence reported in this paper is AF436078.

The positive activator of cephamycin C and clavulanic acid production in Streptomyces clavuligerus is mistranslated in a bldA mutant The GenBank accession number for the sequence reported in this paper is AF436078.

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Evolutionary flux of potentially bldA-dependent Streptomyces genes containing the rare leucine codon TTA

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