Plant and Cell Physiology, 2004, Vol. 45, No. 1 83-91
© 2004 Oxford University Press
Expression and Interaction Analysis of Arabidopsis Skp1-Related Genes
1 Plant Function Exploration Team, Plant Functional Genomics Research Group, Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suyehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045 Japan
2 Department of Biological Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510 Japan
3 Plant Mutation Exploration Team, Plant Functional Genomics Research Group, Genomic Sciences Center, RIKEN Yokohama Institute, 1-7-22 Suyehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045 Japan
4 Plant Molecular Biology Laboratory, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba, Ibaraki, 305-0074 Japan
5 Graduate School of Integrated Science, Yokohama City University, 1-7-29 Suyehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045 Japan
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.
6 Corresponding author: E-mail, minami{at}postman.riken.go.jp; Fax, +81-45-503-9586.
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