Overexpression, purification and characterization of two pyrimidine kinases involved in the biosynthesis of thiamin: 4-amino-5-hydroxymethyl-2-methylpyrimidine kinase and 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate kinase

Reddick, Jason J.; Kinsland, Cynthia; Nicewonger, Robb; Christian, Todd; Downs, Diana M.; Winkler, Malcolm E.; Begley, Tadhg P.

doi:10.1016/s0040-4020(98)01006-0

Cited by 46 publications

(29 citation statements)

References 17 publications

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“…1; Petersen and Downs 1997). The recent demonstration that this enzyme also catalyzes the phosphorylation of 13 makes it likely that thiD is the affected gene in mutants that lack this second activity; therefore, thiD could be the gene previously described as thiJ (Mizote et al 1996;Reddick et al 1998). This latter kinase reaction is likely to be involved in the salvage of 13 from the culture medium and is therefore nonessential for thiamin biosynthesis.…”

Section: Gene Organizationmentioning

confidence: 98%

Thiamin biosynthesis in prokaryotes

Ealick

et al. 1999

Archives of Microbiology

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Twelve genes involved in thiamin biosynthesis in prokaryotes have been identified and overexpressed. Of these, six are required for the thiazole biosynthesis (thiFSGH, thil, and dxs), one is involved in the pyrimidine biosynthesis (thiC), one is required for the linking of the thiazole and the pyrimidine (thiE), and four are kinase genes (thiD, thiM, thiL, and pdxK). The specific reactions catalyzed by ThiEF, Dxs, ThiDM, ThiL, and PdxK have been reconstituted in vitro and ThiS thiocarboxylate has been identified as the sulfur source. The X-ray structures of thiamin phosphate synthase and 5-hydroxyethyl-4-methylthiazole kinase have been completed. The genes coding for the thiamin transport system (thiBPQ) have also been identified. Remaining problems include the cloning and characterization of thiK (thiamin kinase) and the gene(s) involved in the regulation of thiamin biosynthesis. The specific reactions catalyzed by ThiC (pyrimidine formation), and ThiGH and ThiI (thiazole formation) have not yet been identified.

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Section: Gene Organizationmentioning

confidence: 98%

Thiamin biosynthesis in prokaryotes

Ealick

et al. 1999

Archives of Microbiology

Self Cite

270

263

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“…3. Phosphorylations were carried out chemically (21,22) or enzymatically using HMP-P kinase (16,23) or thiamin pyrophosphokinase (13,24,25). HPLC analysis of reaction mixtures containing HMP-PP and either Cof or YmfB showed that both of these enzymes catalyzed the hydrolysis of HMP-PP to HMP-P (Fig.…”

Section: Selection Of Clones Resistant To Bacimethrin or Cf 3 -Hmp-mentioning

confidence: 99%

A Genetic Screen for the Identification of Thiamin Metabolic Genes

Lawhorn

Gerdes

Begley

2004

Journal of Biological Chemistry

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A genetic screen was developed for the identification of genes related to thiamin biosynthesis and degradation. Genes conferring resistance to bacimethrin or 4-amino-2-trifluoromethyl-5-hydroxymethylpyrimidine were selected from Escherichia coli and Bacillus subtilis genomic libraries. Hits from the selection included the known thiamin biosynthetic genes thiC, thiE, and dxs as well as five genes of previously unknown function (E. coli yjjX, yajO, ymfB, and cof and B. subtilis yveN). The gene products YmfB and Cof catalyze the hydrolysis of 4-amino-2-methyl-5-hydroxymethylpyrimidine pyrophosphate to 4-amino-2-methyl-5-hydroxymethylpyrimidine phosphate. YmfB also converts thiamin pyrophosphate into thiamin phosphate.

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“…In bacteria and plants, HMP-PP biosynthesis requires two dedicated enzymes. ThiC produces HMP-P from the purine biosynthetic intermediate 4-aminoimidazole ribotide (1,2), which is then phosphorylated to HMP-PP by ThiD (3,4). In contrast, bacterial THZ-P biosynthesis requires multiple enzymes, and different phyla employ distinct strategies.…”

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confidence: 99%

The Cysteine Desulfhydrase CdsH Is Conditionally Required for Sulfur Mobilization to the Thiamine Thiazole in Salmonella enterica

2014

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Thiamine pyrophosphate is a required coenzyme that contains a mechanistically important sulfur atom. In Salmonella enterica, sulfur is trafficked to both thiamine biosynthesis and 4-thiouridine biosynthesis by the enzyme ThiI using persulfide (R-S-S-H) chemistry. It was previously reported that a thiI mutant strain could grow independent of exogenous thiamine in the presence of cysteine, suggesting there was a second mechanism for sulfur mobilization. Data reported here show that oxidation products of cysteine rescue the growth of a thiI mutant strain by a mechanism that requires the transporter YdjN and the cysteine desulfhydrase CdsH. The data are consistent with a model in which sulfide produced by CdsH reacts with cystine (Cys-S-S-Cys), S-sulfocysteine (Cys-S-SO 3 ؊ ), or another disulfide to form a small-molecule persulfide (R-S-S-H). We suggest that this persulfide replaced ThiI by donating sulfur to the thiamine sulfur carrier protein ThiS. This model describes a potential mechanism used for sulfur trafficking in organisms that lack ThiI but are capable of thiamine biosynthesis.

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Overexpression, purification and characterization of two pyrimidine kinases involved in the biosynthesis of thiamin: 4-amino-5-hydroxymethyl-2-methylpyrimidine kinase and 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate kinase

Cited by 46 publications

References 17 publications

Thiamin biosynthesis in prokaryotes

Thiamin biosynthesis in prokaryotes

A Genetic Screen for the Identification of Thiamin Metabolic Genes

The Cysteine Desulfhydrase CdsH Is Conditionally Required for Sulfur Mobilization to the Thiamine Thiazole in Salmonella enterica

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