Summary: In Streptomyces coelicolor A3(2), two genes, tuf1 and tuf3, encode the apparent polypeptide chain elongation factors EF-Tu1 and EF-Tu3, respectively. While tuf1 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 (Ptuf3) resembles the consensus for the major class of eubacterial promoters, and Ptuf3 was recognized preferentially by an RNA polymerase fraction enriched in αrdB, the principal . factor of S. coelicolor. Nuclease S1 mapping failed to reveal tuf3 transcripts during growth of S. coelicolor in liquid culture, consistent with the apparent absence of EF-Tu3 in total protein extracts of the same strain. However, tuf3 transcription was observed in cultures of S. coelicolor M145 shortly after nutritional shiftdown (which resulted in the disappearance of tuf1 transcripts) and after addition of serine hydroxamate, both of which induce the stringent response. Transcription of tuf3 was also observed in transition-phase and stationary-phase cultures of S. coelicolor J1681, a strain deleted for bldA (which specifies a tRNALeu 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.
AngellS.,
LewisC.G.,
ButtnerM.J.,
BibbM.J.1994; Glucose repression in Streptomyces coelicolor A3(2): a likely regu-latory role for glucose kinase.. Mol & Gen Genet 244:135–143
BrownK.L.,
WoodS.,
ButtnerM.J.1992; Isolation and characterization of the major vegetative RNA polymerase of Streptomyces coelicolor A3(2); renaturation of a sigma subunit using GroEL.. Mol Microbiol 6:1133–1139
ButtnerM.J.,
BrownN.L.1985; RNA polymerase-DNA interactions in Streptomyces. In vitro studies of a S. lividans plasmid promoter with S. coelicolor RNA polymerase.. J Mol Biol 185:177–188
CashelM.,
RuddK.E.1987; The stringent response.. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp. 1410–1438 Edited by
NeidhardtF. C.,
IngrahamJ. L.,
Brooks LowK.,
MagasanikB.,
SchaechterM.,
UmbargerH. E.
Washington, DC: American Society for Microbiology;
ChampnessW.C.,
ChaterK.F.1994; Regulation and integration of antibiotic production and morphological differentiation in Streptomyces spp.. In Regulation of Bacterial Differentiation pp. 61–93 Edited by
PiggotP.,
YoungmanP.,
MoranC.
Washington, DC: American Society for Microbiology;
ChaterK.F.,
BrutonC.J.,
KingA.A.,
SuarezJ.E.1982; The expression of Streptomyces and Escherichia coli drug-resistance determinants cloned into the Streptomyces phage фC31.. Gene 19:21–33
CovarrubiasL.,
BolivarF.1982; Construction and characterization of new cloning vehicles. VI. Plasmid pBR329, a new derivative of pBR328 lacking the 482-base-pair inverted duplication.. Gene 17:79–89
Fernández-MorenoM.A.,
CaballeroJ.L.,
HopwoodD.A.,
MalpartidaF.1991; The act cluster contains regulatory and antibiotic export genes, direct targets for translational control by the hid A transfer RNA gene of Streptomyces.
. Cell 66:769–780
GaalT.,
GourseR.L.1990; Guanosine-3´-diphosphate 5´- diphosphate is not required for growth rate-dependent control of rRNA synthesis in Escherichia coli.
. Proc Natl Acad Sci USA875533–5537
GentryD.R.,
HernandezV.J.,
NguyenL.H.,
JensenD.B.,
CashelM.1993; Synthesis of the stationary-phase sigma factor (δs is positively regulated by ppGpp.. J Bacteriol 175:7982–7989
HernandezV.J.,
BremerH.1990; Guanosine tetraphosphate (ppGpp) dependence of the growth rate control of rrnB PI promoter activity in Escherichia coli.
. J Biol Chem 265:11605–11614
LangeR.,
Hengge-AronisR.1994; The cellular concentration of the (δs subunit of RNA polymerase in Escherichia coli is controlled at the level of transcription, translation, and protein stability.. Genes & Dev 8:1600–1612
LawlorE.J.,
BaylisH.A.,
ChaterK.F.1987; Pleiotropic morphological and antibiotic deficiencies result from mutations in a gene encoding a tRNA-like product in Streptomyces coelicolor A3(2).. Genes & Dev 1:1305–1310
MurrayM.G.1986; Use of sodium trichloroacetate and mung bean nuclease to increase sensitivity and precision during transcript mapping.. Anal Biochem 158:165–170
OchiK.1986; Occurrence of the stringent response in Streptomyces sp. and its significance for the initiation of morphological and physiological differentiation.. J Gen Microbiol 132:2621–2631
OchiK.1987; Metabolic initiation of differentiation and secondary metabolism by Streptomyces griseus: significance of the stringent response (ppGpp) and GTP content in relation to A factor.. J Bacteriol 169:3608–3626
OhlsenK.L.,
GrallaJ.D.1992; Interrelated effects of DNA supercoiling, ppGpp, and low salt on melting within the Escherichia coli rrnB P1 promoter.. Mol Microbiol 6:2243–2251
ReehS.,
PedersenS.,
FriesenJ.D.1976; Biosynthetic regulation of individual proteins in reP and relA strains of Escherichia coli during amino acid starvation.. Mol & Gen Genet 149:279–289
RossW.,
GosinkK.K.,
SalomonJ.,
IgarashiK.,
ZhouC.,
IshihamaA.,
SeverinovK.,
GourseR.L.1993; A third recognition element in bacterial promoters: DNA binding by the subunit of RNA polymerase.. Science 262:1407–1412
RuddK.E.,
BochnerB.R.,
CashelM.,
RothJ.R.1985; Mutations in the spoT gene of Salmonella typhimurium: effects on his operon expression.. J Bacteriol 163:534–542
TakanoE.,
GramajoH.G,
StrauchE.,
AndresN.,
WhiteJ.,
BibbM.J.1992; Transcriptional regulation of the redD trans-criptional activator gene accounts for growth-phase-dependent production of the antibiotic undecylprodigiosin in Streptomyces coelicolor A3(2).. Mol Microbiol 5:289–298
TakayanagiY.,
TanakaK.,
TakahashiH.1994; Structure of the 5´ upstream region and the regulation of the rpoS gene of Escherichia coli.
. Mol & Gen Genet 243:525–531
TanH.,
ChaterK.F.1993; Two developmentally controlled promoters of Streptomyces coelicolor A3(2) that resemble the major class of motility-related promoters in other bacteria.. J Bacteriol 175:933–940
TanakaK.,
TakayanagiY.,
FujitaN.,
IshihamaA.,
TakahashiH.1993; Heterogeneity of the principal σ factor in Escherichia coli-. the rpoS gene product, σ38, is a second principal σ factor of RNA polymerase in stationary-phase Escherichia coli.
. Proc Natl Acad Sci USA903511–3515
VijgenboomE.,
WoudtL.P.,
HeinstraP.W.H.,
RietveldK.,
van HaarlemJ.,
van WezelG.P.,
ShochatS.,
BoschL.1994; Three tuf-like genes in the kirromycin producer Streptomyces ramocissimus.
. Microbiology 140:983–998
van WezelG.P.,
WoudtL.P.,
VervenneR.,
VerdurmenM.L.A.,
VijgenboomE.,
BoschL.1994a; Cloning and sequencing of the tuf genes of Streptomyces coelicolor A3(2).. Biochim Biophys Acta 1219:543–547
van WezelG.P.,
ButtnerM.J.,
VijgenboomE.,
BoschL.,
HopwoodD.A.,
KieserH.M.1995; Mapping of genes involved in macromolecular synthesis on the chromosome of Streptomyces coelicolor A3(2).. J Bacteriol 177:473–476
ZukowskiM.M.,
GaffneyD.F.,
SpeckD.,
KaufmannM.,
FindeliA.,
WisecupA.,
LecocqJ.-P.1983; Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene.. Proc Natl Acad Sci USA801101–1105