Isolation of the ERG2 gene, encoding sterol delta 8-->delta 7 isomerase, from the rice blast fungus Magnaporthe grisea and its expression in the maize smut pathogen Ustilago maydis.
The Magnaporthe grisea ERG2 gene, encoding delta 8-->delta 7 sterol isomerase, was isolated from a genomic library by heterologous hybridization to a fragment of the Ustilago maydis ERG2 gene. The isolated gene contained a reading frame of 745 bp which encoded a protein of 221 amino acids. The coding region was interrupted by a single putative 79-bp-long intron. The deduced amino-acid sequence exhibited similarity to the ERG2 gene products of U. maydis and of Saccharomyces cerevisiae, particularly in the central region of the proteins. The NH2-terminal of all three proteins contained a long stretch of amino acids that were strongly hydrophobic, suggesting that they may function by anchoring the protein to a membrane surface. The M. grisea ERG2 gene complemented a U. maydis deletion mutant in which the ERG2 gene had been removed using a one-step gene replacement procedure. The delta 8-->delta 7 sterol isomerase produced by the M. grisea ERG2 gene exhibited a level of sensitivity to the sterol biosynthesis inhibitor, tridemorph, similar to that of the enzyme derived from the U. maydis ERG2 gene.
Keon
J P
JP
Department of Agricultural Sciences, University of Bristol, Long Ashton Research Station, UK.
James
C S
CS
Court
S
S
Baden-Daintree
C
C
Bailey
A M
AM
Burden
R S
RS
Bard
M
M
Hargreaves
J A
JA
D000595:Amino Acid Sequence; D001483:Base Sequence; D004790:Enzyme Induction; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D005659:Fungicides, Industrial / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D003904:Mitosporic Fungi / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D009025:Morpholines / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013251:Steroid Isomerases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D014170:Transformation, Genetic; D014588:Ustilago / Q000235:genetics*
D005656:Fungal Proteins; D005659:Fungicides, Industrial; D009025:Morpholines; D011993:Recombinant Fusion Proteins; C015554:tridemorph; D013251:Steroid Isomerases; C022669:delta(8)-delta(7)-sterol isomerase