title: Cloning of two splice variants of the rice PTS1 receptor, OsPex5pL and OsPex5pS, and their functional characterization using pex5-deficient yeast and Arabidopsis.
abstract: Using the rice PEX14 cDNA as a bait in a yeast two-hybrid assay, two splice variants of the type I peroxisomal targeting signal (PTS1) receptor, OsPex5pL and OsPex5pS, were cloned from a pathogen-treated rice leaf cDNA library. The proteins were produced from a single gene by alternative splicing, which generated a full-length variant, OsPEX5L, and a variant that lacked exon 7, OsPEX5S. OsPex5pL contained 11 copies of the pentapeptide motif WXXXF/Y in its N-terminus, and seven tetratricopeptide repeats in its C-terminus. Expression of OsPEX5L and OsPEX5S predominantly occurred in leaf tissues, and was induced by various stresses, such as exposure to the pathogen Magnaporthe grisea, and treatment with fungal elicitor, methyl viologen, NaCl or hydrogen peroxide. The Arabidopsis T-DNA insertional pex5 mutant, Atpex5, which does not germinate in the absence of sucrose and was resistant to indole-3-butyric acid (IBA), was perfectly rescued by over-expression of OsPex5pL, but not by OsPex5pS. Using transient expression of OsPex5pL and OsPex5pS in the Atpex5 mutant, we show that OsPex5pL translocates both PTS1- and PTS2-containing proteins into the peroxisome by interacting with OsPex7p, whereas OsPex5pS is involved only in PTS1-dependent import in Arabidopsis.
authors.affiliation: Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea.
authors.forename: Jung Ro
authors.initials: JR
authors.lastname: Lee
authors.forename: Ho Hee
authors.initials: HH
authors.lastname: Jang
authors.forename: Jin Ho
authors.initials: JH
authors.lastname: Park
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Jung
authors.forename: Seung Sik
authors.initials: SS
authors.lastname: Lee
authors.forename: Soo Kwon
authors.initials: SK
authors.lastname: Park
authors.forename: Yong Hun
authors.initials: YH
authors.lastname: Chi
authors.forename: Jeong Chan
authors.initials: JC
authors.lastname: Moon
authors.forename: Young Mee
authors.initials: YM
authors.lastname: Lee
authors.forename: Sun Young
authors.initials: SY
authors.lastname: Kim
authors.forename: Jae-Yean
authors.initials: JY
authors.lastname: Kim
authors.forename: Dae-Jin
authors.initials: DJ
authors.lastname: Yun
authors.forename: Moo Je
authors.initials: MJ
authors.lastname: Cho
authors.forename: Kyun Oh
authors.initials: KO
authors.lastname: Lee
authors.forename: Sang Yeol
authors.initials: SY
authors.lastname: Lee
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D015723:Gene Library; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D000074435:Peroxisome-Targeting Signal 1 Receptor; D020675:Peroxisomes / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D021381:Protein Transport; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D016415:Sequence Alignment; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
chemical_list: D000074435:Peroxisome-Targeting Signal 1 Receptor; D010940:Plant Proteins; D020033:Protein Isoforms; D018160:Receptors, Cytoplasmic and Nuclear