Association of elongation factor 1 alpha and ribosomal protein L3 with the proline-rich region of yeast adenylyl cyclase-associated protein CAP.
CAP is a multifunctional protein; the N-terminal region binds adenylyl cyclase and controls its response to Ras while the C-terminal region is involved in cytoskeletal regulation. In between the two regions, CAP possesses two proline-rich segments, P1 and P2, resembling a consensus sequence for binding SH3 domains. We have identified two yeast proteins with molecular sizes of 48 and 46 kDa associated specifically with P2. Determination of partial protein sequences demonstrated that the 48-kDa and 46-kDa proteins correspond to EF1 alpha and rL3, respectively, neither of which contains any SH3-domain-like sequence. Deletion of P2 from CAP resulted in loss of the activity to bind the two proteins either in vivo or in vitro. Yeast cells whose chromosomal CAP was replaced by the P2-deletion mutant displayed an abnormal phenotype represented by dissociated localizations of CAP and F-actin, which were colocalized in wild-type cells. These results suggest that these associations may have functional significance.
Yanagihara
C
C
Department of Physiology II, Kobe University School of Medicine, Japan.
Shinkai
M
M
Kariya
K
K
Yamawaki-Kataoka
Y
Y
Hu
C D
CD
Masuda
T
T
Kataoka
T
T
D048868:Adaptor Proteins, Signal Transducing; D000262:Adenylyl Cyclases / Q000378:metabolism; D018797:Cell Cycle Proteins; D015825:Chromosomes, Fungal / Q000235:genetics; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D017353:Gene Deletion; D008840:Microfilament Proteins; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000302:isolation & purification / Q000378:metabolism*; D010641:Phenotype; D011392:Proline / Q000378:metabolism*; D011485:Protein Binding; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D048868:Adaptor Proteins, Signal Transducing; C410502:Capt protein, Drosophila; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins; C073751:SRV2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D011392:Proline; D000262:Adenylyl Cyclases