Journal Article

Expression and Interaction Analysis of <i>Arabidopsis Skp1</i>-Related Genes

Naoki Takahashi, Hirofumi Kuroda, Takashi Kuromori, Takashi Hirayama, Motoaki Seki, Kazuo Shinozaki, Hiroaki Shimada and Minami Matsui

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 45, issue 1, pages 83-91
Published in print January 2004 | ISSN: 0032-0781
Published online January 2004 | e-ISSN: 1471-9053 | DOI: https://dx.doi.org/10.1093/pcp/pch009
Expression and Interaction Analysis of Arabidopsis Skp1-Related Genes

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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Specific protein degradation has been observed in several aspects of development and differentiation in many organisms. One example of such proteolysis is regulated by protein polyubiquitination that is promoted by the SCF complex consisting of Skp1, cullin, and an F-box protein. We examined the activities of the Arabidopsis Skp1-related proteins (ASKs). Among 19 annotated ASK genes, we isolated 16 of the corresponding cDNAs (ASK1, 2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19), and examined their gene products for interactions with 24 representatives of F-box proteins carrying various classes of the C-terminal domains using the yeast two-hybrid system. As a result, we found diverse binding specificities: ASK1, ASK2, ASK11 and ASK12 interacted well with COI1, FKF1, UFO-like protein, LRR-containing F-box proteins, and other F-box proteins with unknown C-terminal motifs. We also observed specific interaction between F-box proteins and ASK3, ASK9, ASK13, ASK14, ASK16 and ASK18. In contrast, we detected no interaction between any of the 12 ASK proteins and F-box proteins containing CRFA, CRFB or CRFC domains. Both histochemical and RT-PCR analysis of eight ASK genes expression revealed unique expression patterns for the respective genes.

Keywords: Keywords: Arabidopsis thaliana — ASK — F-box protein — GUS — Protein degradation — Yeast two-hybrid system.; Abbreviations: ASKs, Arabidopsis SKP1-related genes; GUS, β-glucuronidase; RT-PCR, reverse transcription-PCR.

Journal Article.  4962 words.  Illustrated.

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

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