The thiM locus and its relation to phosphorylation of hydroxyethylthiazole in Escherichia coli

Author:

Mizote T1,Nakayama H1

Affiliation:

1. Department of Food and Nutrition, Yamaguchi Women's University, Japan.

Abstract

A mutant of Escherichia coli lacking hydroxyethylthiazole kinase (EC 2.7.1.50) was produced by a further mutation of a temperature-sensitive, auxotrophic mutant for hydroxyethylthiazole. The parent cells possessed two distinct enzymes capable of phosphorylating hydroxyethylthiazole: one was hydroxyethylthiazole kinase, and the other was a phosphotransferase species that required p-nitrophenylphosphate as a phosphoryl donor. Osmotic shock fluid prepared from the mutant cells phosphorylated hydroxyethylthiazole to an extent comparable to that observed with shock fluid from the parent cells, whereas extracts from shocked cells were unable to catalyze the kinase reaction. Shock fluid from a mutant of the other type obtained as a reduced phosphatase activity against p-nitrophenylphosphate did not show any appreciable activity for the phosphotransferase reaction, while extracts from shocked cells showed full kinase activity. The former mutant had lost its ability to grow on hydroxyethylthiazole at high temperature, but the latter mutant still responded to it. It thus appears that the kinase is an enzyme which might play a role in the biosynthesis of thiamine PPi in situ. By conjugation and P1 transduction, a gene governing hydroxyethylthiazole kinase activity, for which we propose the designation thiM, was mapped on the chromosome close to thiD, a gene specifying phosphomethylpyrimidine kinase activity.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

Reference19 articles.

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2. Purification and properties of two acid phosphatase fractions isolated from osmotic shock fluid of Escherichia coli;Dvorak H. F.;Biochemistry,1967

3. Properties of an acid phosphatase in Escherichia coli;Hafkenscheid J. C. M.;Biochim. Biophys. Acta,1968

4. Some recent studies bearing on the one gene-one enzyme hypothesis;Horowitz N. H.;Cold Spring Harbor Symp. Quant. Biol.,1951

5. thiD locus of Escherichia coli;Imamura N.;Experientia,1981

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