The expression of Streptomyces and Escherichist coli drug-resistance determinants cloned into the Streptomyces phage φC31

Chater, Keith F.; Bruton, Celia J.; King, Audrey A.; Súarez, Juan E.

doi:10.1016/0378-1119(82)90185-8

Cited by 120 publications

(38 citation statements)

References 22 publications

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“…S. coelicolor strains M145 (prototrophic, SCP1-SCP2-; Hopwood et al, 1985), Jl50l ( h i d 1 trraA1 strAI Pgl-, SCP1-SCP2-; Chater et al, 1982), J150lAglkA (E. Vijgenboom, unpublished data) and Jl68l (J150lAbldA; Leskiw et al, 1993) were obtained from the John Innes Centre strain collection. E .…”

Section: Methodsmentioning

confidence: 99%

The tuf3 gene of Streptomyces coelicolor A3(2) encodes an inessential elongation factor Tu that is apparently subject to positive stringent control

et al. 1995

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In Streptomyces coelicolor A3(2), two genes, tuf? and tuf3, encode the apparent polypeptide chain elongation factors EF-Tul and EF-Tu3, respectively. While tuff appears to code for the major EF-Tu, the function of tuf3 is unknown. To assess the role of EF-Tu3, tuf3 was subjected to mutational and transcriptional analyses. Replacement of the 5'-half of tuf3 by an antibiotic resistance cassette had no detectable effect on phenotype, indicating that tuf3 is not essential for growth or differentiation. The transcription start site of tuf3 was located approximately 195 nt upstream of the translation start site. The sequence of the tuf3 promoter (PUB) resembles the consensus for the major class of eubacterial promoters, and Pw was recognized preferentially by an RNA polymerase fraction enriched in eB, the principal c factor of 5.coelicolor. Nuclease S1 mapping failed to reveal tuf3 transcripts during growth of 5. coelicolor in liquid culture, consistent with the apparent absence of EFTu3 in total protein extracts of the same strain. However, tuf3 transcription was observed in cultures of 5. coelicolor M145 shortly after nutritional shiftdown (which resulted in the disappearance of tuf? transcripts) and after addition of serine hydroxamate, both of which induce the stringent response. Transcription of tuf3 was also observed in transition-phase and stationaryphase cultures of 5. coelicolor J1681, a strain deleted for bldA (which specifies a t RNA, , for the rare leucine codon UUA). In all of these examples, transcription of tuf3 followed the production of ppGpp, consistent with the hypothesis that tuf3 is subject to positive stringent control.

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Section: Methodsmentioning

confidence: 99%

The tuf3 gene of Streptomyces coelicolor A3(2) encodes an inessential elongation factor Tu that is apparently subject to positive stringent control

et al. 1995

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“…A possible wider occurrence of bldAlike translational regulatory devices is suggested by the discovery that Clostridium acetobutylicum mutants lacking the tRNA for a rare threonine codon are viable but deficient in sporulation and secondary metabolism (43) Bacterial strains, culture conditions, and transformation procedures. S. coelicolor A3(2) strains were J1501 (hisAl uraAl strAl pgl SCP1 SCP2 [10]) and its derivatives J1700 (bld439 [39]) and J1681 (AbldA; this study). Streptomyces lividans 66 TK24 (str-6 SLP2-SLP3- [27]) was the host for promoter-probing studies with pIJ486 and pIJ487 (see below).…”

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confidence: 99%

Accumulation of bldA-specified tRNA is temporally regulated in Streptomyces coelicolor A3(2)

et al. 1993

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Deletion of the bldA gene of Streptomyces coelicolor A3(2), which encodes the only tRNA for the rare UUA codon, had no obvious effects on primary growth but interfered with aerial mycelium formation and antibiotic production. To investigate the possible regulatory role of bldA, its transcription start point was identified, and time courses were determined for the appearance of its primary transcript, the processing of the primary transcript to give a mature 5' end, and the apparent efficiency of translation of ampC mRNA, which contains multiple UUA codons. The bldA promoter was active at all times, but processing of the 5' end of the primary transcript was comparatively inefficient in young cultures. This may perhaps involve an antisense RNA, evidence of which was provided by promoter probing and in vitro transcription. The presence of low levels of the processed form of the tRNA in young cultures followed by increased abundance in older cultures contrasted with the pattern observed for accumulation of a different, presumably typical tRNA which was approximately equally abundant throughout growth. The increased accumulation of the 5' processed form of bldA tRNA coincided with more-efficient translation of ampC mRNA in older cultures, supporting the hypothesis that in at least some physiological conditions, bldA may have a regulatory influence on events late in growth, such as morphological differentiation and antibiotic production.

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“…Streptomyces coelicolor J1501 (hisA1 uraA1 strA1 pgl-1 SCP1 2 , SCP2 2 ; differentiation phenotype as wild-type strain) (Chater et al, 1982) was used as host Abbreviations: CDA, calcium-dependent antibiotic; EMSA, electrophoretic mobility shift assay; 4HPP, 4-hydroxyphenylpyruvate; tsp, transcription start point. strain for gene propagation and gene disruption.…”

Section: Methodsmentioning

confidence: 99%

Autoregulation of hpdR and its effect on CDA biosynthesis in Streptomyces coelicolor

Yang

Wang

et al. 2010

Microbiology

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HpdR, an IclR-family regulator in Streptomyces coelicolor, is a substrate-dependent repressor for the tyrosine catabolic gene hppD. In this study, S1 nuclease protection assays revealed that hpdR is subject to a negative autoregulation. Purified HpdR showed specific DNA-binding activity for the promoter region of hpdR, indicating that the autoregulation of hpdR is performed directly. The disruption of hpdR led to reduced production of CDA by S. coelicolor J1501, suggesting a positive effect of hpdR on CDA biosynthesis. Electrophoretic mobility shift assays showed that HpdR specifically bound to the promoter region of hmaS (SCO3229 in the CDA gene cluster), encoding 4-hydroxymandelic acid synthase. Disruption of hmaS in J1501 abolished CDA production. It is possible that hpdR regulates CDA biosynthesis by controlling the transcription of hmaS.

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The expression of Streptomyces and Escherichist coli drug-resistance determinants cloned into the Streptomyces phage φC31

Cited by 120 publications

References 22 publications

The tuf3 gene of Streptomyces coelicolor A3(2) encodes an inessential elongation factor Tu that is apparently subject to positive stringent control

The tuf3 gene of Streptomyces coelicolor A3(2) encodes an inessential elongation factor Tu that is apparently subject to positive stringent control

Accumulation of bldA-specified tRNA is temporally regulated in Streptomyces coelicolor A3(2)

Autoregulation of hpdR and its effect on CDA biosynthesis in Streptomyces coelicolor

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