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. 1993 Jun;175(12):3905–3908. doi: 10.1128/jb.175.12.3905-3908.1993

Transcriptional analysis of the gap-pgk-tpi-ppc gene cluster of Corynebacterium glutamicum.

J W Schwinde 1, N Thum-Schmitz 1, B J Eikmanns 1, H Sahm 1
PMCID: PMC204809  PMID: 7685337

Abstract

The transcriptional organization of the Corynebacterium glutamicum gap-pgk-tpi-ppc gene cluster, encoding glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, triosephosphate isomerase, and phosphoenolpyruvate carboxylase, was investigated by Northern (RNA) blot and primer extension analyses. Four transcripts corresponding to gap, to gap-pgk-tpi, to pgk-tpi, and to pgk-tpi-ppc were identified. The respective transcriptional initiation sites in front of gap and pgk were located, and, from the analysis of DNA sequences upstream of these and of previously determined transcriptional start sites, common structures which may be important for promoter function in C. glutamicum are discussed.

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Selected References

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  1. Alefounder P. R., Perham R. N. Identification, molecular cloning and sequence analysis of a gene cluster encoding the class II fructose 1,6-bisphosphate aldolase, 3-phosphoglycerate kinase and a putative second glyceraldehyde 3-phosphate dehydrogenase of Escherichia coli. Mol Microbiol. 1989 Jun;3(6):723–732. doi: 10.1111/j.1365-2958.1989.tb00221.x. [DOI] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowen D., Littlechild J. A., Fothergill J. E., Watson H. C., Hall L. Nucleotide sequence of the phosphoglycerate kinase gene from the extreme thermophile Thermus thermophilus. Comparison of the deduced amino acid sequence with that of the mesophilic yeast phosphoglycerate kinase. Biochem J. 1988 Sep 1;254(2):509–517. doi: 10.1042/bj2540509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Branlant C., Oster T., Branlant G. Nucleotide sequence determination of the DNA region coding for Bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase and of the flanking DNA regions required for its expression in Escherichia coli. Gene. 1989 Jan 30;75(1):145–155. doi: 10.1016/0378-1119(89)90391-0. [DOI] [PubMed] [Google Scholar]
  5. Börmann E. R., Eikmanns B. J., Sahm H. Molecular analysis of the Corynebacterium glutamicum gdh gene encoding glutamate dehydrogenase. Mol Microbiol. 1992 Feb;6(3):317–326. doi: 10.1111/j.1365-2958.1992.tb01474.x. [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. Conway T., Ingram L. O. Phosphoglycerate kinase gene from Zymomonas mobilis: cloning, sequencing, and localization within the gap operon. J Bacteriol. 1988 Apr;170(4):1926–1933. doi: 10.1128/jb.170.4.1926-1933.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies G. J., Littlechild J. A., Watson H. C., Hall L. Sequence and expression of the gene encoding 3-phosphoglycerate kinase from Bacillus stearothermophilus. Gene. 1991 Dec 20;109(1):39–45. doi: 10.1016/0378-1119(91)90586-z. [DOI] [PubMed] [Google Scholar]
  9. Eddy C. K., Mejia J. P., Conway T., Ingram L. O. Differential expression of gap and pgk genes within the gap operon of Zymomonas mobilis. J Bacteriol. 1989 Dec;171(12):6549–6554. doi: 10.1128/jb.171.12.6549-6554.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eikmanns B. J., Follettie M. T., Griot M. U., Sinskey A. J. The phosphoenolpyruvate carboxylase gene of Corynebacterium glutamicum: molecular cloning, nucleotide sequence, and expression. Mol Gen Genet. 1989 Aug;218(2):330–339. doi: 10.1007/BF00331286. [DOI] [PubMed] [Google Scholar]
  11. Eikmanns B. J. Identification, sequence analysis, and expression of a Corynebacterium glutamicum gene cluster encoding the three glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, and triosephosphate isomerase. J Bacteriol. 1992 Oct;174(19):6076–6086. doi: 10.1128/jb.174.19.6076-6086.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eikmanns B. J., Kleinertz E., Liebl W., Sahm H. A family of Corynebacterium glutamicum/Escherichia coli shuttle vectors for cloning, controlled gene expression, and promoter probing. Gene. 1991 Jun 15;102(1):93–98. doi: 10.1016/0378-1119(91)90545-m. [DOI] [PubMed] [Google Scholar]
  13. Han K. S., Archer J. A., Sinskey A. J. The molecular structure of the Corynebacterium glutamicum threonine synthase gene. Mol Microbiol. 1990 Oct;4(10):1693–1702. doi: 10.1111/j.1365-2958.1990.tb00546.x. [DOI] [PubMed] [Google Scholar]
  14. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Krug M. S., Berger S. L. First-strand cDNA synthesis primed with oligo(dT). Methods Enzymol. 1987;152:316–325. doi: 10.1016/0076-6879(87)52036-5. [DOI] [PubMed] [Google Scholar]
  16. Marcel T., Archer J. A., Mengin-Lecreulx D., Sinskey A. J. Nucleotide sequence and organization of the upstream region of the Corynebacterium glutamicum lysA gene. Mol Microbiol. 1990 Nov;4(11):1819–1830. doi: 10.1111/j.1365-2958.1990.tb02030.x. [DOI] [PubMed] [Google Scholar]
  17. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  19. O'Regan M., Thierbach G., Bachmann B., Villeval D., Lepage P., Viret J. F., Lemoine Y. Cloning and nucleotide sequence of the phosphoenolpyruvate carboxylase-coding gene of Corynebacterium glutamicum ATCC13032. Gene. 1989 Apr 30;77(2):237–251. doi: 10.1016/0378-1119(89)90072-3. [DOI] [PubMed] [Google Scholar]
  20. Peoples O. P., Liebl W., Bodis M., Maeng P. J., Follettie M. T., Archer J. A., Sinskey A. J. Nucleotide sequence and fine structural analysis of the Corynebacterium glutamicum hom-thrB operon. Mol Microbiol. 1988 Jan;2(1):63–72. doi: 10.1111/j.1365-2958.1988.tb00007.x. [DOI] [PubMed] [Google Scholar]
  21. Schläpfer B. S., Zuber H. Cloning and sequencing of the genes encoding glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and triosephosphate isomerase (gap operon) from mesophilic Bacillus megaterium: comparison with corresponding sequences from thermophilic Bacillus stearothermophilus. Gene. 1992 Dec 1;122(1):53–62. doi: 10.1016/0378-1119(92)90031-j. [DOI] [PubMed] [Google Scholar]
  22. von der Osten C. H., Barbas C. F., 3rd, Wong C. H., Sinskey A. J. Molecular cloning, nucleotide sequence and fine-structural analysis of the Corynebacterium glutamicum fda gene: structural comparison of C. glutamicum fructose-1,6-biphosphate aldolase to class I and class II aldolases. Mol Microbiol. 1989 Nov;3(11):1625–1637. doi: 10.1111/j.1365-2958.1989.tb00148.x. [DOI] [PubMed] [Google Scholar]

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