A chemogenomic screening of sulfanilamide-hypersensitive Saccharomyces cerevisiae mutants uncovers ABZ2, the gene encoding a fungal aminodeoxychorismate lyase.
Large-scale phenotypic analyses have proved to be useful strategies in providing functional clues about the uncharacterized yeast genes. We used here a chemogenomic profiling of yeast deletion collections to identify the core of cellular processes challenged by treatment with the p-aminobenzoate/folate antimetabolite sulfanilamide. In addition to sulfanilamide-hypersensitive mutants whose deleted genes can be categorized into a number of groups, including one-carbon related metabolism, vacuole biogenesis and vesicular transport, DNA metabolic and cell cycle processes, and lipid and amino acid metabolism, two uncharacterized open reading frames (YHI9 and YMR289w) were also identified. A detailed characterization of YMR289w revealed that this gene was required for growth in media lacking p-aminobenzoic or folic acid and encoded a 4-amino-4-deoxychorismate lyase, which is the last of the three enzymatic activities required for p-aminobenzoic acid biosynthesis. In light of these results, YMR289w was designated ABZ2, in accordance with the accepted nomenclature. ABZ2 was able to rescue the p-aminobenzoate auxotrophy of an Escherichia coli pabC mutant, thus demonstrating that ABZ2 and pabC are functional homologues. Phylogenetic analyses revealed that Abz2p is the founder member of a new group of fungal 4-amino-4-deoxychorismate lyases that have no significant homology to its bacterial or plant counterparts. Abz2p appeared to form homodimers and dimerization was indispensable for its catalytic activity.
Botet
Javier
J
Departamento de Microbiología y Genética, Instituto de Microbiología Bioquímica, Universidad de Salamanca/CSIC, Campus Miguel de Unamuno, 37007 Salamanca, Spain.
Mateos
Laura
L
Revuelta
José L
JL
Santos
María A
MA
D010129:4-Aminobenzoic Acid / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism; D019281:Dimerization; D004926:Escherichia coli; D005492:Folic Acid / Q000378:metabolism; D005800:Genes, Fungal; D023281:Genomics / Q000379:methods*; D008190:Lyases / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D007652:Oxo-Acid-Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010944:Plants / Q000201:enzymology; D011387:Prokaryotic Cells / Q000201:enzymology; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity / Q000187:drug effects; D000077145:Sulfanilamide; D013424:Sulfanilamides / Q000494:pharmacology*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D013424:Sulfanilamides; C079404:PabC protein, Bacteria; D000077145:Sulfanilamide; D005492:Folic Acid; D008190:Lyases; D007652:Oxo-Acid-Lyases; C066600:aminodeoxychorismate lyase; D010129:4-Aminobenzoic Acid