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J. Biol. Chem., Vol. 279, Issue 42, 43555-43559, October 15, 2004

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A Genetic Screen for the Identification of Thiamin Metabolic Genes^*

Brian G. Lawhorn, Svetlana Y. Gerdes, and Tadhg P. Begley¶

From the Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853 and Integrated Genomics, Inc., Chicago, Illinois 60612

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.

Received for publication, April 19, 2004 , and in revised form, July 19, 2004.

^* This work was supported by National Institutes of Health Grant DK44083. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains additional Experimental Procedures.

^¶ To whom correspondence should be addressed. Tel.: 607-255-7133; E-mail: tpb2{at}cornell.edu.

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