Abstract
Pantothenate kinase catalyzes the rate-controlling step in coenzyme A (CoA) biosynthesis. The structural gene (coaA) located at 90 min of the Escherichia coli chromosome was cloned and sequenced. The coaA gene was transcribed in the opposite direction to the flanking genes birA and thrU and produced a single 1.1-kb transcript. Translation of the coaA gene produced two protein products (36.4 and 35.4 kDa) that differed by eight amino acids at the amino terminus. The poor homology of the coaA promoter region to consensus E. coli promoter sequences and the low frequency of optimal codon usage (0.565) were consistent with the low abundance of pantothenate kinase. Strains containing multiple copies of the coaA gene possessed 76-fold-higher specific activity of pantothenate kinase; however, there was only a 2.7-fold increase in the steady-state level of CoA. These data corroborate the conclusion that regulation of pantothenate kinase activity by feedback inhibition is the critical factor controlling the intracellular CoA concentration.
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- Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
- An G., Friesen J. D. The nucleotide sequence of tufB and four nearby tRNA structural genes of Escherichia coli. Gene. 1980 Dec;12(1-2):33–39. doi: 10.1016/0378-1119(80)90013-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Chin D. T., Goff S. A., Webster T., Smith T., Goldberg A. L. Sequence of the lon gene in Escherichia coli. A heat-shock gene which encodes the ATP-dependent protease La. J Biol Chem. 1988 Aug 25;263(24):11718–11728. [PubMed] [Google Scholar]
- Flaks J. G., Leboy P. S., Birge E. A., Kurland C. G. Mutations and genetics concerned with the ribosome. Cold Spring Harb Symp Quant Biol. 1966;31:623–631. doi: 10.1101/sqb.1966.031.01.081. [DOI] [PubMed] [Google Scholar]
- Henikoff S. Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 1987;155:156–165. doi: 10.1016/0076-6879(87)55014-5. [DOI] [PubMed] [Google Scholar]
- Howard P. K., Shaw J., Otsuka A. J. Nucleotide sequence of the birA gene encoding the biotin operon repressor and biotin holoenzyme synthetase functions of Escherichia coli. Gene. 1985;35(3):321–331. doi: 10.1016/0378-1119(85)90011-3. [DOI] [PubMed] [Google Scholar]
- Ikemura T. Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes: a proposal for a synonymous codon choice that is optimal for the E. coli translational system. J Mol Biol. 1981 Sep 25;151(3):389–409. doi: 10.1016/0022-2836(81)90003-6. [DOI] [PubMed] [Google Scholar]
- Jackowski S., Alix J. H. Cloning, sequence, and expression of the pantothenate permease (panF) gene of Escherichia coli. J Bacteriol. 1990 Jul;172(7):3842–3848. doi: 10.1128/jb.172.7.3842-3848.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Altered molecular form of acyl carrier protein associated with beta-ketoacyl-acyl carrier protein synthase II (fabF) mutants. J Bacteriol. 1987 Apr;169(4):1469–1473. doi: 10.1128/jb.169.4.1469-1473.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Metabolism of 4'-phosphopantetheine in Escherichia coli. J Bacteriol. 1984 Apr;158(1):115–120. doi: 10.1128/jb.158.1.115-120.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Ratio of active to inactive forms of acyl carrier protein in Escherichia coli. J Biol Chem. 1983 Dec 25;258(24):15186–15191. [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Regulation of coenzyme A biosynthesis. J Bacteriol. 1981 Dec;148(3):926–932. doi: 10.1128/jb.148.3.926-932.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackowski S., Rock C. O. Turnover of the 4'-phosphopantetheine prosthetic group of acyl carrier protein. J Biol Chem. 1984 Feb 10;259(3):1891–1895. [PubMed] [Google Scholar]
- Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Platt T. Transcription termination and the regulation of gene expression. Annu Rev Biochem. 1986;55:339–372. doi: 10.1146/annurev.bi.55.070186.002011. [DOI] [PubMed] [Google Scholar]
- 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]
- Showe M. K., DeMoss J. A. Localization and regulation of synthesis of nitrate reductase in Escherichia coli. J Bacteriol. 1968 Apr;95(4):1305–1313. doi: 10.1128/jb.95.4.1305-1313.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Song W. J., Jackowski S. coaA and rts are allelic and located at kilobase 3532 on the Escherichia coli physical map. J Bacteriol. 1992 Mar;174(5):1705–1706. doi: 10.1128/jb.174.5.1705-1706.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
- Vallari D. S., Jackowski S. Biosynthesis and degradation both contribute to the regulation of coenzyme A content in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3961–3966. doi: 10.1128/jb.170.9.3961-3966.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallari D. S., Jackowski S., Rock C. O. Regulation of pantothenate kinase by coenzyme A and its thioesters. J Biol Chem. 1987 Feb 25;262(6):2468–2471. [PubMed] [Google Scholar]
- Vallari D. S., Rock C. O. Isolation and characterization of temperature-sensitive pantothenate kinase (coaA) mutants of Escherichia coli. J Bacteriol. 1987 Dec;169(12):5795–5800. doi: 10.1128/jb.169.12.5795-5800.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]