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. 1989 Oct;171(10):5567–5571. doi: 10.1128/jb.171.10.5567-5571.1989

(S)-3-hydroxy-3-methylglutaryl coenzyme A reductase, a product of the mva operon of Pseudomonas mevalonii, is regulated at the transcriptional level.

Y L Wang 1, M J Beach 1, V W Rodwell 1
PMCID: PMC210398  PMID: 2477360

Abstract

We have cloned and sequenced a 505-base-pair (bp) segment of DNA situated upstream of mvaA, the structural gene for (S)-3-hydroxy-3-methylglutaryl coenzyme A reductase (EC 1.1.1.88) of Pseudomonas mevalonii. The DNA segment that we characterized includes the promoter region for the mva operon. Nuclease S1 mapping and primer extension analysis showed that mvaA is the promoter-proximal gene of the mva operon. Transcription initiates at -56 bp relative to the first A (+1) of the translation start site. Transcription in vivo was induced by mevalonate. Structural features of the mva promoter region include an 80-bp A + T-rich region, and -12, -24 consensus sequences that resemble sequences of sigma 54 promoters in enteric organisms. The relative amplitudes of catalytic activity, enzyme protein, and mvaA mRNA are consistent with a model of regulation of this operon at the transcriptional level.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beach M. J., Rodwell V. W. Cloning, sequencing, and overexpression of mvaA, which encodes Pseudomonas mevalonii 3-hydroxy-3-methylglutaryl coenzyme A reductase. J Bacteriol. 1989 Jun;171(6):2994–3001. doi: 10.1128/jb.171.6.2994-3001.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Buck M. Deletion analysis of the Klebsiella pneumoniae nitrogenase promoter: importance of spacing between conserved sequences around positions -12 and -24 for activation by the nifA and ntrC (glnG) products. J Bacteriol. 1986 May;166(2):545–551. doi: 10.1128/jb.166.2.545-551.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  6. Calzone F. J., Britten R. J., Davidson E. H. Mapping of gene transcripts by nuclease protection assays and cDNA primer extension. Methods Enzymol. 1987;152:611–632. doi: 10.1016/0076-6879(87)52069-9. [DOI] [PubMed] [Google Scholar]
  7. Cole S. T., Bremer E., Hindennach I., Henning U. Characterisation of the promoters for the ompA gene which encodes a major outer membrane protein of Escherichia coli. Mol Gen Genet. 1982;188(3):472–479. doi: 10.1007/BF00330051. [DOI] [PubMed] [Google Scholar]
  8. Cortay J. C., Rieul C., Duclos B., Cozzone A. J. Characterization of the phosphoproteins of Escherichia coli cells by electrophoretic analysis. Eur J Biochem. 1986 Sep 1;159(2):227–237. doi: 10.1111/j.1432-1033.1986.tb09858.x. [DOI] [PubMed] [Google Scholar]
  9. Cozzone A. J. Protein phosphorylation in prokaryotes. Annu Rev Microbiol. 1988;42:97–125. doi: 10.1146/annurev.mi.42.100188.000525. [DOI] [PubMed] [Google Scholar]
  10. Dixon R. The xylABC promoter from the Pseudomonas putida TOL plasmid is activated by nitrogen regulatory genes in Escherichia coli. Mol Gen Genet. 1986 Apr;203(1):129–136. doi: 10.1007/BF00330393. [DOI] [PubMed] [Google Scholar]
  11. Ebina Y., Nakazawa A. Cyclic AMP-dependent initiation and rho-dependent termination of colicin E1 gene transcription. J Biol Chem. 1983 Jun 10;258(11):7072–7078. [PubMed] [Google Scholar]
  12. Gill J. F., Jr, Beach M. J., Rodwell V. W. Mevalonate utilization in Pseudomonas sp. M. Purification and characterization of an inducible 3-hydroxy-3-methylglutaryl coenzyme A reductase. J Biol Chem. 1985 Aug 5;260(16):9393–9398. [PubMed] [Google Scholar]
  13. Gill J. F., Jr, Beach M. J., Rodwell V. W. Transport of mevalonate by Pseudomonas sp. strain M. J Bacteriol. 1984 Oct;160(1):294–298. doi: 10.1128/jb.160.1.294-298.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hirschman J., Wong P. K., Sei K., Keener J., Kustu S. Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7525–7529. doi: 10.1073/pnas.82.22.7525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hu L., Phillips A. T. Organization and multiple regulation of histidine utilization genes in Pseudomonas putida. J Bacteriol. 1988 Sep;170(9):4272–4279. doi: 10.1128/jb.170.9.4272-4279.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Inouye S., Ebina Y., Nakazawa A., Nakazawa T. Nucleotide sequence surrounding transcription initiation site of xylABC operon on TOL plasmid of Pseudomonas putida. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1688–1691. doi: 10.1073/pnas.81.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inouye S., Nakazawa A., Nakazawa T. Nucleotide sequence of the regulatory gene xylS on the Pseudomonas putida TOL plasmid and identification of the protein product. Gene. 1986;44(2-3):235–242. doi: 10.1016/0378-1119(86)90187-3. [DOI] [PubMed] [Google Scholar]
  18. Jaurin B., Cohen S. N. Streptomyces contain Escherichia coli-type A + T-rich promoters having novel structural features. Gene. 1985;39(2-3):191–201. doi: 10.1016/0378-1119(85)90313-0. [DOI] [PubMed] [Google Scholar]
  19. Johnson K., Parker M. L., Lory S. Nucleotide sequence and transcriptional initiation site of two Pseudomonas aeruginosa pilin genes. J Biol Chem. 1986 Nov 25;261(33):15703–15708. [PubMed] [Google Scholar]
  20. Kennelly P. J., Rodwell V. W. Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase by reversible phosphorylation-dephosphorylation. J Lipid Res. 1985 Aug;26(8):903–914. [PubMed] [Google Scholar]
  21. Khan H., Buck M., Dixon R. Deletion loop mutagenesis of the nifL promoter from Klebsiella pneumoniae: role of the -26 to -12 region in promoter function. Gene. 1986;45(3):281–288. doi: 10.1016/0378-1119(86)90026-0. [DOI] [PubMed] [Google Scholar]
  22. Klumpp D. J., Plank D. W., Bowdin L. J., Stueland C. S., Chung T., LaPorte D. C. Nucleotide sequence of aceK, the gene encoding isocitrate dehydrogenase kinase/phosphatase. J Bacteriol. 1988 Jun;170(6):2763–2769. doi: 10.1128/jb.170.6.2763-2769.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. LaPorte D. C., Koshland D. E., Jr Phosphorylation of isocitrate dehydrogenase as a demonstration of enhanced sensitivity in covalent regulation. Nature. 1983 Sep 22;305(5932):286–290. doi: 10.1038/305286a0. [DOI] [PubMed] [Google Scholar]
  24. Lin E. S., Wilson D. B. Transcription of the celE gene in Thermomonospora fusca. J Bacteriol. 1988 Sep;170(9):3838–3842. doi: 10.1128/jb.170.9.3838-3842.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mermod N., Lehrbach P. R., Reineke W., Timmis K. N. Transcription of the TOL plasmid toluate catabolic pathway operon of Pseudomonas putida is determined by a pair of co-ordinately and positively regulated overlapping promoters. EMBO J. 1984 Nov;3(11):2461–2466. doi: 10.1002/j.1460-2075.1984.tb02156.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Merrick M. J., Gibbins J. R. The nucleotide sequence of the nitrogen-regulation gene ntrA of Klebsiella pneumoniae and comparison with conserved features in bacterial RNA polymerase sigma factors. Nucleic Acids Res. 1985 Nov 11;13(21):7607–7620. doi: 10.1093/nar/13.21.7607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  28. Minton N. P., Clarke L. E. Identification of the promoter of the Pseudomonas gene coding for carboxypeptidase G2. J Mol Appl Genet. 1985;3(1):26–35. [PubMed] [Google Scholar]
  29. Musso R. E., Di Lauro R., Adhya S., de Crombrugghe B. Dual control for transcription of the galactose operon by cyclic AMP and its receptor protein at two interspersed promoters. Cell. 1977 Nov;12(3):847–854. doi: 10.1016/0092-8674(77)90283-5. [DOI] [PubMed] [Google Scholar]
  30. Nesin M., Lupski J. R., Godson G. N. Role of the 5' upstream sequence and tandem promoters in regulation of the rpsU-dnaG-rpoD macromolecular synthesis operon. J Bacteriol. 1988 Dec;170(12):5759–5764. doi: 10.1128/jb.170.12.5759-5764.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reitzer L. J., Magasanik B. Expression of glnA in Escherichia coli is regulated at tandem promoters. Proc Natl Acad Sci U S A. 1985 Apr;82(7):1979–1983. doi: 10.1073/pnas.82.7.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rodwell V. W., Nordstrom J. L., Mitschelen J. J. Regulation of HMG-CoA reductase. Adv Lipid Res. 1976;14:1–74. doi: 10.1016/b978-0-12-024914-5.50008-5. [DOI] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Song C. S., Yu J. H., Bai D. H., Hester P. Y., Kim K. H. Antibodies to the alpha-subunit of insulin receptor from eggs of immunized hens. J Immunol. 1985 Nov;135(5):3354–3359. [PubMed] [Google Scholar]
  35. Stephens R. S., Wagar E. A., Edman U. Developmental regulation of tandem promoters for the major outer membrane protein gene of Chlamydia trachomatis. J Bacteriol. 1988 Feb;170(2):744–750. doi: 10.1128/jb.170.2.744-750.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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