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Cloning of two splice variants of the rice PTS1 receptor, Os Pex5pL and Os Pex5pS, and their functional characterization using pex5 ‐deficient yeast and Arabidopsis
Author(s) -
Lee Jung Ro,
Jang Ho Hee,
Park Jin Ho,
Jung Ji Hyun,
Lee Seung Sik,
Park Soo Kwon,
Chi Yong Hun,
Moon Jeong Chan,
Lee Young Mee,
Kim Sun Young,
Kim JaeYean,
Yun DaeJin,
Cho Moo Je,
Lee Kyun Oh,
Lee Sang Yeol
Publication year - 2006
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2006.02797.x
Subject(s) - arabidopsis , cloning (programming) , splice , yeast , biology , receptor , genetics , computational biology , microbiology and biotechnology , gene , computer science , mutant , programming language
Summary 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, Os Pex5pL and Os Pex5pS, 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 . Os Pex5pL 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 Os Pex5pL, but not by Os Pex5pS. Using transient expression of Os Pex5pL and Os Pex5pS in the Atpex5 mutant, we show that Os Pex5pL translocates both PTS1‐ and PTS2‐containing proteins into the peroxisome by interacting with Os Pex7p, whereas Os Pex5pS is involved only in PTS1‐dependent import in Arabidopsis.