Abstract
Two chromosomal loci containing the Corynebacterium glutamicum ATCC 17965 proB and proC genes were isolated by complementation of Escherichia coli proB and proC auxotrophic mutants. Together with a proA gene described earlier, these new genes describe the major C. glutamicum proline biosynthetic pathway. The proB and proA genes, closely linked in most bacteria, are in C. glutamicum separated by a 304-amino-acid open reading frame (unk) whose predicted sequence resembles that of the 2-hydroxy acid dehydrogenases. C. glutamicum mutants that carry null alleles of proB, proA, and proC were constructed or isolated from mutagenized cultures. Single proC mutants are auxotrophic for proline and secrete delta1-pyrroline-5-carboxylate, which are the expected phenotypes of bacterial proC mutants. However, the phenotypes or proB and proA mutants are unexpected. A proB mutant has a pleiotropic phenotype, being both proline auxotrophic and affected in cell morphology. Null proA alleles still grow slowly under proline starvation, which suggests that a proA-independent bypass of this metabolic step exists in C. glutamicum. Since proA mutants are complemented by a plasmid that contains the wild-type asd gene of C. glutamicum, the asd gene may play a role in this bypass.
Full Text
The Full Text of this article is available as a PDF (272.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adams E., Frank L. Metabolism of proline and the hydroxyprolines. Annu Rev Biochem. 1980;49:1005–1061. doi: 10.1146/annurev.bi.49.070180.005041. [DOI] [PubMed] [Google Scholar]
- Ankri S., Reyes O., Leblon G. Electrotransformation of highly DNA-restrictive corynebacteria with synthetic DNA. Plasmid. 1996 Jan;35(1):62–66. doi: 10.1006/plas.1996.0007. [DOI] [PubMed] [Google Scholar]
- Arst H. N., Jr, Jones S. A., Bailey C. R. A method for the selection of deletion mutations in the L-proline catabolism gene cluster of Aspergillus nidulans. Genet Res. 1981 Oct;38(2):171–195. doi: 10.1017/s0016672300020516. [DOI] [PubMed] [Google Scholar]
- Arthur M., Molinas C., Dutka-Malen S., Courvalin P. Structural relationship between the vancomycin resistance protein VanH and 2-hydroxycarboxylic acid dehydrogenases. Gene. 1991 Jul 15;103(1):133–134. doi: 10.1016/0378-1119(91)90405-z. [DOI] [PubMed] [Google Scholar]
- Berg C. M., Rossi J. J. Proline excretion and indirect suppression in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1974 Jun;118(3):928–934. doi: 10.1128/jb.118.3.928-934.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dandekar A. M., Uratsu S. L. A single base pair change in proline biosynthesis genes causes osmotic stress tolerance. J Bacteriol. 1988 Dec;170(12):5943–5945. doi: 10.1128/jb.170.12.5943-5945.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Follettie M. T., Peoples O. P., Agoropoulou C., Sinskey A. J. Gene structure and expression of the Corynebacterium flavum N13 ask-asd operon. J Bacteriol. 1993 Jul;175(13):4096–4103. doi: 10.1128/jb.175.13.4096-4103.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fons M., Cami B., Patte J. C., Chippaux M. Cloning in Escherichia coli of genes involved in the synthesis of proline and leucine in Desulfovibrio desulfuricans Norway. Mol Gen Genet. 1987 Jan;206(1):141–143. doi: 10.1007/BF00326549. [DOI] [PubMed] [Google Scholar]
- Gowrishankar J. Identification of osmoresponsive genes in Escherichia coli: evidence for participation of potassium and proline transport systems in osmoregulation. J Bacteriol. 1985 Oct;164(1):434–445. doi: 10.1128/jb.164.1.434-445.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayzer D. J., Leisinger T. Proline biosynthesis in Escherichia coli. Purification and characterisation of glutamate-semialdehyde dehydrogenase. Eur J Biochem. 1982 Jan;121(3):561–565. doi: 10.1111/j.1432-1033.1982.tb05823.x. [DOI] [PubMed] [Google Scholar]
- Heath J. D., Weinstock G. M. Tandem duplications of the lac region of the Escherichia coli chromosome. Biochimie. 1991 Apr;73(4):343–352. doi: 10.1016/0300-9084(91)90099-m. [DOI] [PubMed] [Google Scholar]
- Henikoff S., Henikoff J. G. Automated assembly of protein blocks for database searching. Nucleic Acids Res. 1991 Dec 11;19(23):6565–6572. doi: 10.1093/nar/19.23.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Labarre J., Reyes O., Guyonvarch A., Leblon G. Gene replacement, integration, and amplification at the gdhA locus of Corynebacterium glutamicum. J Bacteriol. 1993 Feb;175(4):1001–1007. doi: 10.1128/jb.175.4.1001-1007.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Louie H., Chan V. L. Cloning and characterization of the gamma-glutamyl phosphate reductase gene of Campylobacter jejuni. Mol Gen Genet. 1993 Jul;240(1):29–35. doi: 10.1007/BF00276880. [DOI] [PubMed] [Google Scholar]
- Marck C. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res. 1988 Mar 11;16(5):1829–1836. doi: 10.1093/nar/16.5.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ogura M., Kawata-Mukai M., Itaya M., Takio K., Tanaka T. Multiple copies of the proB gene enhance degS-dependent extracellular protease production in Bacillus subtilis. J Bacteriol. 1994 Sep;176(18):5673–5680. doi: 10.1128/jb.176.18.5673-5680.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reyes O., Guyonvarch A., Bonamy C., Salti V., David F., Leblon G. 'Integron'-bearing vectors: a method suitable for stable chromosomal integration in highly restrictive corynebacteria. Gene. 1991 Oct 30;107(1):61–68. doi: 10.1016/0378-1119(91)90297-o. [DOI] [PubMed] [Google Scholar]
- Rose R. E. The nucleotide sequence of pACYC184. Nucleic Acids Res. 1988 Jan 11;16(1):355–355. doi: 10.1093/nar/16.1.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
- Serebrijski I., Reyes O., Leblon G. Corrected gene assignments of Escherichia coli pro- mutations. J Bacteriol. 1995 Dec;177(24):7261–7264. doi: 10.1128/jb.177.24.7261-7264.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Serebrijski I., Wojcik F., Reyes O., Leblon G. Multicopy suppression by asd gene and osmotic stress-dependent complementation by heterologous proA in proA mutants. J Bacteriol. 1995 Dec;177(24):7255–7260. doi: 10.1128/jb.177.24.7255-7260.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith C. J., Deutch A. H., Rushlow K. E. Purification and characteristics of a gamma-glutamyl kinase involved in Escherichia coli proline biosynthesis. J Bacteriol. 1984 Feb;157(2):545–551. doi: 10.1128/jb.157.2.545-551.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whatmore A. M., Chudek J. A., Reed R. H. The effects of osmotic upshock on the intracellular solute pools of Bacillus subtilis. J Gen Microbiol. 1990 Dec;136(12):2527–2535. doi: 10.1099/00221287-136-12-2527. [DOI] [PubMed] [Google Scholar]
- 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]
