Expression and interaction analysis of Arabidopsis Skp1-related genes.
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.
Takahashi
Naoki
N
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.
Kuroda
Hirofumi
H
Kuromori
Takashi
T
Hirayama
Takashi
T
Seki
Motoaki
M
Shinozaki
Kazuo
K
Shimada
Hiroaki
H
Matsui
Minami
M
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D044783:F-Box Proteins / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D018745:Genome, Plant; D005810:Multigene Family / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010802:Phylogeny; D030821:Plants, Genetically Modified / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D044786:S-Phase Kinase-Associated Proteins / Q000235:genetics* / Q000378:metabolism
C120363:ASK1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D044783:F-Box Proteins; D044786:S-Phase Kinase-Associated Proteins