Identification of Escherichia coli ubiB , a Gene Required for the First Monooxygenase Step in Ubiquinone Biosynthesis

Abstract

ABSTRACT It was recently discovered that the aarF gene in Providencia stuartii is required for coenzyme Q (CoQ) biosynthesis. Here we report that yigR , the Escherichia coli homologue of aarF , is ubiB , a gene required for the first monooxygenase step in CoQ biosynthesis. Both the P. stuartii aarF and E. coli ubiB ( yigR ) disruption mutant strains lack CoQ and accumulate octaprenylphenol. Octaprenylphenol is the CoQ biosynthetic intermediate found to accumulate in the E. coli strain AN59, which contains the ubiB409 mutant allele. Analysis of the mutation in the E. coli strain AN59 reveals no mutations within the ubiB gene, but instead shows the presence of an IS 1 element at position +516 of the ubiE gene. The ubiE gene encodes a C -methyltransferase required for the synthesis of both CoQ and menaquinone, and it is the 5′ gene in an operon containing ubiE , yigP , and ubiB . The data indicate that octaprenylphenol accumulates in AN59 as a result of a polar effect of the ubiE ::IS 1 mutation on the downstream ubiB gene. AN59 is complemented by a DNA segment containing the contiguous ubiE , yigP , and ubiB genes. Although transformation of AN59 with a DNA segment containing the ubiB coding region fails to restore CoQ biosynthesis, transformation with the ubiE coding region results in a low-frequency but significant rescue attributed to homologous recombination. In addition, the fre gene, previously considered to correspond to ubiB , was found not to be involved in CoQ biosynthesis. The ubiB gene is a member of a predicted protein kinase family of which the Saccharomyces cerevisiae ABC1 gene is the prototypic member. The possible protein kinase function of UbiB and Abc1 and the role these polypeptides may play in CoQ biosynthesis are discussed.