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A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase.
J Biol Chem. 1985 Dec 05; 260(28):15291-7.JB

Abstract

The gene which codes for isocitrate dehydrogenase kinase/phosphatase of Escherichia coli, aceK, has been cloned. Physical and functional mapping of this clone indicated that both the isocitrate dehydrogenase kinase and isocitrate dehydrogenase phosphatase activities are encoded by an 1800-base pair sequence. This sequence produced a polypeptide with an apparent molecular weight of 66,000, which is identical to that of the purified protein. Since a protein of this size would require an 1800-base pair coding sequence, we conclude that isocitrate dehydrogenase kinase and isocitrate dehydrogenase phosphatase are expressed from a single gene. This strongly suggests that both activities reside on the same polypeptide chain. The cloning of aceK was made possible by the fortuitous addition of a second origin of replication to the expression vectors which were employed. These expression vectors were found to inhibit the growth of E. coli on the minimal acetate selective medium. The inclusion of a second origin of replication reduced the copy number and so reduced the inhibitory effects of these vectors. Control of the copy number through the addition of replication origins may have a general facility when manipulating plasmids which are potentially toxic to E. coli.

Authors

No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

2999109

Citation

LaPorte, D C., and T Chung. "A Single Gene Codes for the Kinase and Phosphatase Which Regulate Isocitrate Dehydrogenase." The Journal of Biological Chemistry, vol. 260, no. 28, 1985, pp. 15291-7.
LaPorte DC, Chung T. A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase. J Biol Chem. 1985;260(28):15291-7.
LaPorte, D. C., & Chung, T. (1985). A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase. The Journal of Biological Chemistry, 260(28), 15291-7.
LaPorte DC, Chung T. A Single Gene Codes for the Kinase and Phosphatase Which Regulate Isocitrate Dehydrogenase. J Biol Chem. 1985 Dec 5;260(28):15291-7. PubMed PMID: 2999109.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A single gene codes for the kinase and phosphatase which regulate isocitrate dehydrogenase. AU - LaPorte,D C, AU - Chung,T, PY - 1985/12/5/pubmed PY - 1985/12/5/medline PY - 1985/12/5/entrez SP - 15291 EP - 7 JF - The Journal of biological chemistry JO - J Biol Chem VL - 260 IS - 28 N2 - The gene which codes for isocitrate dehydrogenase kinase/phosphatase of Escherichia coli, aceK, has been cloned. Physical and functional mapping of this clone indicated that both the isocitrate dehydrogenase kinase and isocitrate dehydrogenase phosphatase activities are encoded by an 1800-base pair sequence. This sequence produced a polypeptide with an apparent molecular weight of 66,000, which is identical to that of the purified protein. Since a protein of this size would require an 1800-base pair coding sequence, we conclude that isocitrate dehydrogenase kinase and isocitrate dehydrogenase phosphatase are expressed from a single gene. This strongly suggests that both activities reside on the same polypeptide chain. The cloning of aceK was made possible by the fortuitous addition of a second origin of replication to the expression vectors which were employed. These expression vectors were found to inhibit the growth of E. coli on the minimal acetate selective medium. The inclusion of a second origin of replication reduced the copy number and so reduced the inhibitory effects of these vectors. Control of the copy number through the addition of replication origins may have a general facility when manipulating plasmids which are potentially toxic to E. coli. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/2999109/A_single_gene_codes_for_the_kinase_and_phosphatase_which_regulate_isocitrate_dehydrogenase_ DB - PRIME DP - Unbound Medicine ER -