title: Identification and characterization of Candida utilis multidrug efflux transporter CuCdr1p.
abstract: The edible, nitrate assimilating, yeast Candida utilis is a commercial food additive, and it is a potentially useful host for heterologous protein expression. A number of ATP-binding cassette (ABC) transporters are multidrug efflux pumps that can cause multidrug resistance in opportunistic pathogens. In order to develop optimal novel antimicrobial agents it is imperative to understand the structure, function and expression of these transporters. With the ultimate aim of developing an alternative yeast host for the heterologous expression of eukaryotic membrane transporters, and to identify ABC transporters potentially associated with C. utilis multidrug resistance, we classified the entire repertoire of 30 C. utilis ABC proteins. We named the open reading frame most similar to the archetype multidrug efflux pump gene C. albicans CDR1 as CuCDR1 Overexpression of CuCDR1 in Saccharomyces cerevisiae ADΔ caused multidrug resistance similar to that of cells overexpressing CaCDR1 Unlike CaCdr1p, however, the C-terminally green fluorescent protein (GFP) tagged CuCdr1p-GFP was functionally impaired and did not properly localize to the plasma membrane. CuCdr1p function could be recovered however by adding a 15 amino acid linker -GAGGSAGGSGGAGAG- between CuCdr1p and the C-terminal GFP tag.
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-J3-7, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors.forename: Wittawan
authors.initials: W
authors.lastname: Watanasrisin
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-J3-7, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors.forename: Shun
authors.initials: S
authors.lastname: Iwatani
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-J3-7, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Oura
authors.affiliation: Central Laboratories for Key Technologies, KIRIN Company, Ltd, 1-13-5 Fukuura, Kanazawa-ku Yokohama 236-0004, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Tomita
authors.affiliation: Central Laboratories for Key Technologies, KIRIN Company, Ltd, 1-13-5 Fukuura, Kanazawa-ku Yokohama 236-0004, Japan.
authors.forename: Shigehiro
authors.initials: S
authors.lastname: Ikushima
authors.affiliation: Mycology Unit, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, 1873 Rama4, Pathumwan, Wangmai, Bangkok 10330, Thailand.
authors.forename: Ariya
authors.initials: A
authors.lastname: Chindamporn
authors.affiliation: Mycology Unit, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, 1873 Rama4, Pathumwan, Wangmai, Bangkok 10330, Thailand Department of Oral Sciences, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, 310 Great King Street, Dunedin 9054, New Zealand.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Niimi
authors.affiliation: Department of Oral Sciences, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, 310 Great King Street, Dunedin 9054, New Zealand.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Niimi
authors.affiliation: Department of Oral Sciences, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, 310 Great King Street, Dunedin 9054, New Zealand.
authors.forename: Erwin
authors.initials: E
authors.lastname: Lamping
authors.affiliation: Department of Oral Sciences, Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, 310 Great King Street, Dunedin 9054, New Zealand.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cannon
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-J3-7, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan skajiwar@bio.titech.ac.jp.
authors.forename: Susumu
authors.initials: S
authors.lastname: Kajiwara
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism*; D000935:Antifungal Agents / Q000494:pharmacology; D002175:Candida / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D026141:Drug Resistance, Multiple, Fungal; D015870:Gene Expression; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D011994:Recombinant Proteins
keywords: ABC transporters; ATP-binding cassette (ABC) proteins; Candida utilis; heterologous expression; multidrug efflux pump; multidrug resistance; tandem gene duplication