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Microbiology 142 (1996), 2515-2523; DOI  10.1099/00221287-142-9-2515
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Isolation of the mRNA-capping enzyme and ferric-reductase-related genes from Candida albicans

Toshiko Yamada-Okabe1,4, Osamu Shimmi2, Rikuo Doi1, Kiyohisa Mizumoto3, Mikio Arisawa2 and Hisafumi Yamada-Okabe2

Department of Hygiene, School of Medicine, Yokohama City University, Japan
Department of Mycology, Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa 247, Japan
Department of Physiological Chemistry, School of Pharmaceutical Science, Kitasato University, Japan

4Author for correspondence: Hisafumi Yamada-Okabe. Tel: +81 467 47 2213. Fax: +81 467 46 5320. e-mail: hisafumi.okabe@roche.com

ABSTRACT

The mRNA-capping enzyme (mRNA 5'-guanylyltransferase) gene was cloned from a Candida albicans genomic DNA library by functional complementation of a Saccharomyces cerevisiae ceg1{Delta} null mutation. This gene, referred to as CGT1 (C. albicans guanylyltransferase 1), can encode a 52 kDa protein that is highly homologous to S. cerevisiae Ceg1p. CGT1 in a single-copy plasmid complemented the lethality of the S. cerevisiae ceg1{Delta} null mutation and, like S. cerevisiae Ceg1p, bacterially expressed Cgt1p was able to form a stable complex with the GMP moiety of GTP and to synthesize the cap structure in vitro, demonstrating that CGT1 is the C. albicans mRNA 5'-guanylyltransferase gene. CGT1 seemed to exist as a single copy in the C. albicans genome and was actively transcribed into mRNA. Another ORF was found in an opposite strand very close to the CGT1 locus. This gene shared significant sequence homology with S. cerevisiae FRE1, the gene encoding ferric reductase, and therefore was designated CFL1 (C. albicans ferric-reductase-like gene 1). Despite its sequence homology with S. cerevisiae FRE1, CFL1 mRNA was not induced by iron deprivation, and CFL1 did not complement the slow growth of a S. cerevisiae fre1{Delta} null mutant in the absence of iron, suggesting that CFL1 is functionally distinct from S. cerevisiae FRE1.

Keywords: mRNA-capping enzyme, guanylyltransferase, Candida albicans, stable GMP-enzyme complex, ferric reductase




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