Journal Article

Direct Interaction and Determination of Binding Domains among Peroxisomal Import Factors in <i>Arabidopsis thaliana</i>

Kazumasa Nito, Makoto Hayashi and Mikio Nishimura

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 4, pages 355-366
Published in print April 2002 | ISSN: 0032-0781
Published online April 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf057
Direct Interaction and Determination of Binding Domains among Peroxisomal Import Factors in Arabidopsis thaliana

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We analyzed the role of Arabidopsis orthologues of human Pex14p, Pex5p and Pex7p that are central components of peroxisomal protein import machinery. Immunoblot analysis showed that AtPex14p and AtPex5p were present in most organs in Arabidopsis, suggesting that these factors play a role in the main protein import pathways for plant peroxisomes. Two-hybrid analysis showed that AtPex14p interacted with AtPex5p, but not with AtPex7p. In addition, AtPex7p was bound to AtPex5p, indicating that the PTS2 pathway depends on the PTS1 pathway in Arabidopsis. Further analysis showed that the nine WXXXF/Y repeats in the amino acids 231K-450D and 1M-230V of AtPex5p are bound to two N-terminal domains, amino acids 58I-65L and 78R-97R of AtPex14p and the C-terminal amino acids 266Y-317S of AtPex7p, respectively. Since the binding domains of AtPex5p to AtPex14p and AtPex7p do not overlap, AtPex14p, AtPex5p and AtPex7p might form their complex and function cooperatively in peroxisomal protein import.

Keywords: Key words: Arabidopsis — Peroxin — Peroxisome — Protein import — Protein–protein interaction.; Abbreviation: GFP, green fluorescent protein; GST, glutathione S-transferase; HPR, hydroxypyruvate reductase; ICL, isocitrate lyase; IgG, immunoglobulin G; PTS, peroxisome targeting signal; TPR, tetratricopeptide repeat; Trx, thioredoxin.

Journal Article.  8381 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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