Identification of a CAP (adenylyl-cyclase-associated protein) homologous gene in Lentinus edodes and its functional complementation of yeast CAP mutants

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Abstract
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 mushroomLentinus edodes. TheL. edodes capgene contains eight introns and an ORF encoding a 518 amino acid protein. TheL. edodesCAP is 35.5% and 40.9% identical at the amino acid level withSaccharomyces cerevisiaeCAP andSchizosaccharomyces pombeCAP, 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 thatL. edodes capis a sinμle-copy gene and uniformly expressed. Expression of theL. edodesCAP in bothSchiz. pombeandSacch.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 theL. edodesCAP andSchiz. pombeactin. This result and the functional complementation test indicate that CAP fromL. edodeshas a conserved C-terminal domain function.