Streptomycespromoter-probe plasmids that utilise thexyIEgene ofPseudomonas putida

Clayton, Timothy M.; Bibb, Mervyn J.

doi:10.1093/nar/18.4.1077

Cited by 71 publications

(41 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transformants were streaked on solid SLAB medium, and after 2 to 4 days of growth at 30ЊC, they were sprayed with a 0.5 M catechol solution and inspected for the yellow color of hydroxymuconic semialdehyde (8,9). Positive colonies were grown in 50 ml of SLAB liquid medium at 30ЊC on a rotary shaker.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Crude extracts were obtained after ultrasonication (Branson sonifier) and removal of cell debris by centrifugation (20,000 ϫ g, 20 min). Aliquots were tested with the substrate catechol as described before (8,9).DNA isolation and manipulations. Total and plasmid DNAs were isolated from Streptomyces strains as described by Hopwood et al (16).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Minimal requirements of the Streptomyces lividans 66 oriC region and its transcriptional and translational activities

Zakrzewska‐Czerwińska

Majka

Schrempf

1995

J Bacteriol

View full text Add to dashboard Cite

Deletion analysis of a previously constructed minichromosome revealed that a stretch of DNA which is longer than 623 bp but shorter than 837 bp is required for autonomous replication of the Streptomyces lividans chromosome. Each of the dnaA and dnaN genes flanking the oriC region is individually transcribed from two promoters. Within the intergenic, nontranslatable region between the dnaA and dnaN genes, five main transcripts and several less abundant transcripts of various lengths as well as one of the promoters were identified. The introduction of additional DnaA boxes in S. lividans led to a significant increase in dnaA gene transcripts and to an enhanced level of the DnaA (73-kDa) protein. In summary, the data suggest that dnaA gene transcription is autoregulated and that initiation of the S. lividans chromosome is tightly controlled.The replication of several bacterial chromosomes was found to be initiated at the origin of replication, oriC. The features of oriC and, with a few exceptions, the arrangement of genes in its vicinity are remarkably similar among eubacteria (25, 38). Sequence analyses revealed that the origins of various eubacteria contain highly conserved segments which are separated by variable spacer regions. The DnaA box motifs (TTATC/AC AC/AA) (36) constituting the binding sites for the initiator protein DnaA are prominent in the conserved parts of oriC regions from various bacteria. The only relevant variation is an A-to-G exchange at the third position of the DnaA box motif within GC-rich DNA present in gram-positive bacteria such as Micrococcus luteus (10) and Streptomyces lividans (42). Most eubacteria contain a block of genes, dnaA-dnaN-recF-gyrB (10, 38), encoding the initiator protein DnaA, the ␤-subunit of DNA polymerase III holoenzyme, a product for recombination, and the ␤-subunit of DNA gyrase, respectively. Within most bacteria, oriC is situated close to the dnaA gene. In Escherichia coli, the arrangement of the dnaA region matches that in other bacteria; however, this region is located about 42 kb away from oriC (35).Procaryotic chromosomal replication has been best studied in E. coli. Its DnaA protein is a basic protein of 52 kDa to which ATP binds. About 10 to 40 ATP-DnaA molecules interact with four DnaA boxes within oriC (19,36). According to a current model (14), the newly synthesized DnaA protein is titrated by these DnaA boxes. As soon as the number of DnaA molecules exceeds the number of boxes, initiation is supposed to occur. A high number of GATC sites which can be methylated by the Dam methyltransferase have been found in the oriC region of E. coli, as well as in other members of the family Enterobacteriaceae (44), and GANTC sites are present in the Caulobacter crescentus oriC (43).Contrary to many bacteria, Streptomyces species grow as substrate mycelia which differentiate to aerial mycelia and spores upon depletion of nutrients. Depending on the species, the size of the GC-rich Streptomyces chromosome ranges between 6.6 and 8 Mb (18,32). Recently, we identified the...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Minimal requirements of the Streptomyces lividans 66 oriC region and its transcriptional and translational activities

Zakrzewska‐Czerwińska

Majka

Schrempf

1995

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…All the promoters were sequenced using pUC/M13 forward and reverse primers in a Perkin Elmer ABI PRISM 310 Genetic Analyzer, extracted from the plasmid via digestion with HindIII/BglII or HindIII/BamHI and ligated into pIJ4083 in order to construct transcriptional fusions with the xylE reporter gene (Clayton & Bibb, 1990). The obtained promoter-probe plasmids pIJ4083-PphoA, pIJ4083-PphoC and pIJ4083-PphoD were first transformed into Streptomyces lividans, then extracted, confirmed by restriction with HindIII and XbaI and transformed into S. coelicolor.…”

Section: A K Apel and Othersmentioning

confidence: 99%

Phosphate control of phoA, phoC and phoD gene expression in Streptomyces coelicolor reveals significant differences in binding of PhoP to their promoter regions

et al. 2007

View full text Add to dashboard Cite

Three putative alkaline phosphatase genes, phoA, phoC and phoD, were identified in the genome of Streptomyces coelicolor by homology with the amino acid sequence obtained from the PhoA protein of Streptomyces griseus. PhoA and PhoC correspond to broad-spectrum alkaline phosphatases whereas PhoD is similar to a Ca 2+ -dependent phospholipase D of Streptomyces chromofuscus. The phoA and phoD genes were efficiently expressed in R5 medium under phosphate-limited conditions, as shown by studies using the xylE reporter gene, whereas phoC was poorly transcribed under the same conditions. Expression of phoA was clearly PhoPdependent since it was not transcribed in the S. coelicolor DphoP mutant and was strongly activated under low phosphate concentrations. Similarly, expression of phoD was PhoPdependent and highly sensitive to phosphate availability. By contrast, expression of phoC was not PhoP-dependent. Electrophoretic mobility shift assays showed that PhoP binds to the phoA and phoD promoters, but not to that of phoC. Footprinting studies with GST-PhoP revealed the presence of a PHO box (two direct 11 nt repeats) in the phoA promoter and two PHO boxes in the promoter of phoD. The transcription start points of the three promoters were identified by primer extension. The transcription start point of phoD coincides with the G of its translation start codon, indicating that this gene is transcribed as a leaderless mRNA. The deduced "10 and "35 regions of phoD (but not those of phoA) overlapped with the PHO boxes in this promoter, suggesting that an excess of PhoP interferes with binding of the RNA polymerase to this promoter. In summary, the three promoters showed clear differences in the modulation of their expression by PhoP.

show abstract

“…15 Primers were designed (Supplementary Table S1) to amplify the regions upstream of thnQ, thnP, thnK, thnI and thnH (Figure 1a). Amplification products for the corresponding fragments, obtained with Pfx polymerase (Invitrogen S.A. El Prat De Llobregat, Barcelona, Spain), were cloned into pIJ4083.…”

Section: Regulation Of Thienamycin Biosynthesis In Streptomyces Cattleyamentioning

confidence: 99%

Transcriptional organization of ThnI-regulated thienamycin biosynthetic genes in Streptomyces cattleya

et al. 2010

View full text Add to dashboard Cite

Streptomycespromoter-probe plasmids that utilise thexyIEgene ofPseudomonas putida

Cited by 71 publications

References 4 publications

Minimal requirements of the Streptomyces lividans 66 oriC region and its transcriptional and translational activities

Minimal requirements of the Streptomyces lividans 66 oriC region and its transcriptional and translational activities

Phosphate control of phoA, phoC and phoD gene expression in Streptomyces coelicolor reveals significant differences in binding of PhoP to their promoter regions

Transcriptional organization of ThnI-regulated thienamycin biosynthetic genes in Streptomyces cattleya

Contact Info

Product

Resources

About