title: Genetic evidence for involvement of membrane trafficking in the action of 5-fluorouracil.
abstract: To identify novel genes that mediate cellular sensitivity and resistance to 5-fluorouracil (5-FU), we performed a genome-wide genetic screening to identify altered susceptibility to 5-FU by Schizosaccharomyces pombe haploid nonessential gene deletion library containing 3004 deletion mutants. We identified 50 hypersensitive and 12 resistant mutants to this drug. Mutants sensitive or resistant to 5-FU were classified into various categories based on their putative functions. The largest group of the genes whose disruption renders cells altered susceptibility to 5-FU is involved in nucleic acid metabolism, but to our surprise, the second largest group is involved in membrane trafficking. In addition, several other membrane traffic mutants examined including gdi1-i11, ypt3-i5, Δryh1, Δric1, and Δaps1 exhibited hypersensitivity to 5-FU. Furthermore, we found that 5-FU in low concentration that generally do not affect cell growth altered the localization of Syb1, a secretory vesicle SNARE synaptobrevin which is cycled between the plasma membrane and the endocytic pathway. Notably, 5-FU at such low concentration also significantly inhibited the secretion of acid phosphatase. Altogether, our findings revealed the first evidence that 5-FU influences membrane trafficking as the potential underlying mechanism of the drug action.
authors.affiliation: Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province 110112, China; Division of Pharmacology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
authors.forename: Lingling
authors.initials: L
authors.lastname: Hu
authors.affiliation: Department of Breast Surgery and Surgical Oncology, Research Unit of General Surgery, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning Province 110001, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yao
authors.affiliation: Division of Pharmacology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
authors.forename: Yan
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province 110112, China.
authors.forename: Qiannan
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province 110112, China.
authors.forename: Si
authors.initials: S
authors.lastname: Chen
authors.affiliation: Division of Pharmacology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Hayafuji
authors.affiliation: Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province 110112, China; Division of Pharmacology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Kuno
authors.affiliation: Department of Microbial and Biochemical Pharmacy, School of Pharmacy, China Medical University, No. 77 Puhe Road, Shenyang North New Area, Shenyang, Liaoning Province 110112, China. Electronic address: fangyue@mail.cmu.edu.cn.
authors.forename: Yue
authors.initials: Y
authors.lastname: Fang
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D005472:Fluorouracil / Q000494:pharmacology*; D017353:Gene Deletion; D016681:Genome, Fungal / Q000187:drug effects; D006238:Haploidy; D009696:Nucleic Acids / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000235:genetics; D012568:Schizosaccharomyces / Q000187:drug effects / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D009696:Nucleic Acids; D029702:Schizosaccharomyces pombe Proteins; D005472:Fluorouracil
keywords: 5-Fluorouracil; Fission yeast; Membrane trafficking