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PMID:8432721

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Citation

Vander Horn, PB, Backstrom, AD, Stewart, V and Begley, TP (1993) Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in Escherichia coli K-12. J. Bacteriol. 175:982-92

Abstract

Escherichia coli K-12 synthesizes thiamine pyrophosphate (vitamin B1) de novo. Two precursors [4-methyl-5-(beta-hydroxyethyl)thiazole monophosphate and 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate] are coupled to form thiamine monophosphate, which is then phosphorylated to make thiamine pyrophosphate. Previous studies have identified two classes of thi mutations, clustered at 90 min on the genetic map, which result in requirements for the thiazole or the hydroxymethylpryimidine. We report here our initial molecular genetic analysis of the thi cluster. We cloned the thi cluster genes and examined their organization, structure, and function by a combination of phenotypic testing, complementation analysis, polypeptide expression, and DNA sequencing. We found five tightly linked genes, designated thiCEFGH. The thiC gene product is required for the synthesis of the hydroxymethylpyrimidine. The thiE, thiF, thiG, and thiH gene products are required for synthesis of the thiazole. These mutants did not respond to 1-deoxy-D-threo-2-pentulose, indicating that they are blocked in the conversion of this precursor compound to the thiazole itself.

Links

PubMed PMC193010

Keywords

Amino Acid Sequence; Base Sequence; Chromosome Mapping; Cloning, Molecular; DNA, Bacterial/genetics; Escherichia coli/genetics; Escherichia coli Proteins; Genes, Bacterial; Genetic Complementation Test; Molecular Sequence Data; Nucleotidyltransferases/genetics; Restriction Mapping; Sequence Alignment; Thiamine/biosynthesis

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:THIF

acts_upstream_of_or_within

GO:0052837: thiazole biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:THIG

acts_upstream_of_or_within

GO:0009228: thiamine biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:THIH

acts_upstream_of_or_within

GO:0009228: thiamine biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:THIE

GO:0009228: thiamine biosynthetic process

ECO:0000315:

P

Figure 3

complete
CACAO 4984

ECOLI:THIG

involved_in

GO:0009228: thiamine biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:THIH

involved_in

GO:0009228: thiamine biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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