Identification of a CAP (adenylyl-cyclase-associated protein) homologous gene in Lentinus edodes and its functional complementation of yeast CAP mutants.
The adenylyl-cyclase-associated protein, CAP, was originally identified in yeasts as a protein that functions in both signal transduction and cytoskeletal organization. This paper reports the identification of a cDNA and genomic DNA that encodes a CAP homologue from the mushroom Lentinus edodes. The L. edodes cap gene contains eight introns and an ORF encoding a 518 amino acid protein. The L. edodes CAP is 35.5% and 40.9% identical at the amino acid level with Saccharomyces cerevisiae CAP and Schizosaccharomyces pombe CAP, respectively. The C-terminal domain shows greater homology (39-46% identity) with yeast CAPs than does the N-terminal domain (27-35% identity). Southern blotting and Northern blotting results suggest that L. edodes cap is a single-copy gene and uniformly expressed. Expression of the L. edodes CAP in both Schiz. pombe and Sacch. cerevisiae complemented defects associated with the loss of the C-terminal domain function of the endogenous CAP. By using a yeast two-hybrid assay, an interaction was demonstrated between the L. edodes CAP and Schiz. pombe actin. This result and the functional complementation test indicate that CAP from L. edodes has a conserved C-terminal domain function.
Zhou
Guo-Lei
GL
1 Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue 690, Japan.
Miyazaki
Yasumasa
Y
2 Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226, Japan.
Nakagawa
Tsuyoshi
T
3 Research Institute of Molecular Genetics, Shimane University, Matsue 690, Japan.
Tanaka
Katsunori
K
1 Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue 690, Japan.
Shishido
Kazuo
K
2 Department of Life Science, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226, Japan.
Matsuda
Hideyuki
H
1 Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue 690, Japan.
Kawamukai
Makoto
M
1 Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue 690, Japan.
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D001483:Base Sequence; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary / Q000096:biosynthesis; D004271:DNA, Fungal / Q000096:biosynthesis; D029721:Drosophila Proteins; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics*; D005816:Genetic Complementation Test; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D010641:Phenotype; D010802:Phylogeny; D011124:Polyporaceae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment
D048868:Adaptor Proteins, Signal Transducing; C410502:Capt protein, Drosophila; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D004271:DNA, Fungal; D029721:Drosophila Proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; C073751:SRV2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins