TY - JOUR
T1 - Expression and Interaction Analysis of Arabidopsis Skp1-Related Genes
AU - Takahashi, Naoki
AU - Kuroda, Hirofumi
AU - Kuromori, Takashi
AU - Hirayama, Takashi
AU - Seki, Motoaki
AU - Shinozaki, Kazuo
AU - Shimada, Hiroaki
AU - Matsui, Minami
N1 - Funding Information:
We thank Dr. Takashi Ito (Kanazawa University) for providing us with the mating type plasmid vectors and yeast strains, and Dr. Tsuyoshi Nakagawa (Shimane University) for providing us with binary vectors. We thank ABRC and the Salk Institute Genomic Analysis Laboratory for providing the sequence-indexed Arabidopsis T-DNA insertion mutants. We thank Ms. Yasue Ichikawa and Rie Nakazawa for DNA sequencing (Bioarchitect Research Group, RIKEN). This work was supported in part by a Grant-in-Aid for Young Scientists (B) from the Ministry of Education, Culture, Sports, and Technology (14740448 to H.K.).
PY - 2004/1
Y1 - 2004/1
N2 - 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.
AB - 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.
KW - ASK
KW - Arabidopsis thaliana
KW - F-box protein
KW - GUS
KW - Protein degradation
KW - Yeast two-hybrid system
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U2 - 10.1093/pcp/pch009
DO - 10.1093/pcp/pch009
M3 - Article
C2 - 14749489
AN - SCOPUS:1042268311
VL - 45
SP - 83
EP - 91
JO - Plant and Cell Physiology
JF - Plant and Cell Physiology
SN - 0032-0781
IS - 1
ER -