The spacer protein Spc110p targets calmodulin to the central plaque of the yeast spindle pole body.
Yeast calmodulin (CaM) was found to be localized to the microtubule organizing centre, the spindle pole body. The spindle pole body is a multi-layered structure consisting of outer, central and inner plaques. In this paper, we report that a fraction of CaM is in association with the central plaque of the spindle pole body. This localization is dependent on the calmodulin-binding site of another spindle pole body component, Spc110p, which serves as a spacer connecting the inner plaque with the central plaque. Since the CaM-binding site of Spc110p is located near the carboxy terminus, Spc110p-dependent localization of calmodulin defines the orientation of Spc110p with the carboxy terminus towards the central plaque and the amino terminus towards the inner plaque. This orientation of Spc110p was confirmed using antibodies specific for the amino-terminal end of Spc110p, which predominantly labelled the inner plaque. In addition, synthetic peptides corresponding to the calmodulin-binding site of Spc110p bound to calmodulin with a Kd in the nanomolar range and nearly independent of Ca2+.
Spang
A
A
Max-Planck Institut für Biochemie, Genzentrum, Martinsried, Germany.
Grein
K
K
Schiebel
E
E
D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins; D018385:Centrosome / Q000378:metabolism* / Q000648:ultrastructure; D003598:Cytoskeletal Proteins; D017931:DNA Primers / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D005656:Fungal Proteins; D009687:Nuclear Proteins; D010455:Peptides; C073277:SPC110 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D002118:Calcium