Construction and functional analysis of fatty acid desaturase gene disruptants in Candida albicans.
Polyunsaturated fatty acids (PUFAs), including linoleic acid (C18 : 2) and alpha-linolenic acid (C18 : 3), are major components of membranes. PUFAs are produced from monounsaturated fatty acids by several fatty acid desaturases (FADs) in many fungi, but Saccharomyces cerevisiae, Schizosaccharomyces pombe and humans do not have these enzymes. Although the fungal pathogen Candida albicans produces C18 : 2 and C18 : 3, the enzymes that synthesize them have not yet been investigated. In this report, two ORFs, CaFAD2 and CaFAD3, were identified based on their homology to other yeast FADs, and CaFAD2 and CaFAD3 gene disruptants were constructed. Cafad2Delta and Cafad3Delta lost their ability to produce C18 : 2 and C18 : 3, respectively. Furthermore, S. cerevisiae cells expressing CaFad2p converted palmitoleic acid (C16 : 1) and C18 : 1 to hexadecadienoic acid (C16 : 2) and C18 : 2, respectively, and CaFad3p-expressing cells converted C18 : 2 to C18 : 3. These results strongly supported that CaFAD2 encodes the Delta12 FAD and that CaFAD3 encodes the omega3 FAD. However, phenotypic analysis demonstrated that the presence of these PUFAs did not affect the virulence to mice, or morphogenesis in the culture media used to induce morphological change of C. albicans.
Murayama
Somay Yamagata
SY
Laboratory of Infectious Agents Surveillance, Kitasato Institute for Life Sciences and Graduate School of Infection Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Negishi
Yumiko
Y
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.|Department of Bioactive Molecules, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.|Laboratory of Infectious Agents Surveillance, Kitasato Institute for Life Sciences and Graduate School of Infection Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Umeyama
Takashi
T
Department of Bioactive Molecules, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Kaneko
Aki
A
Department of Bioactive Molecules, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Oura
Takahiro
T
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Niimi
Masakazu
M
Department of Bioactive Molecules, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Ubukata
Kimiko
K
Laboratory of Infectious Agents Surveillance, Kitasato Institute for Life Sciences and Graduate School of Infection Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Kajiwara
Susumu
S
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
D000595:Amino Acid Sequence; D000818:Animals; D002176:Candida albicans / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D002177:Candidiasis / Q000382:microbiology; D004195:Disease Models, Animal; D044943:Fatty Acid Desaturases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D005227:Fatty Acids / Q000032:analysis; D005229:Fatty Acids, Monounsaturated / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D019787:Linoleic Acid / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014774:Virulence; D017962:alpha-Linolenic Acid / Q000096:biosynthesis
D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D011994:Recombinant Proteins; D017962:alpha-Linolenic Acid; C008757:palmitoleic acid; D019787:Linoleic Acid; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase; C078565:omega-3 fatty acid desaturase