Spc110p: assembly properties and role in the connection of nuclear microtubules to the yeast spindle pole body.
Spc110p is an essential component of the budding yeast spindle pole body (SPB). It binds calmodulin and contains a long central coiled-coil rod which acts as a spacer element between the central plaque of the SPB and the ends of the nuclear or spindle microtubules. This suggests that the essential function of Spc110p is to connect the nuclear microtubules to the SPB. To confirm this, we examined the phenotype of ts alleles of SPC110, one of which contains a mutation in the calmodulin binding site and was suppressed by overexpression of calmodulin. The alleles fail to form a functional mitotic spindle because spindle microtubules are not properly connected to the SPB. We also examined the phenotype of the toxic overexpression of either the wild-type or a truncated version of Spc110p containing a deletion of most of the coiled-coil domain. Both of these proteins form large ordered spheroidal polymers in the nucleus. The polymerization of the truncated Spc110p appears to be initiated inside the SPB from the position where Spc110p is normally located, and as the polymer grows in size it severs the connection between the nuclear microtubules and the SPB. The polymers were purified and are composed of Spc110p and calmodulin. A model for the structure of the polymer is proposed.
MRC Laboratory of Molecular Biology, Cambridge, UK.
J V
JV
Kilmartin
P Y
PY
Goh
D001665:Binding Sites; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins; D002467:Cell Nucleus / Q000378:metabolism; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008870:Microtubules / Q000378:metabolism*; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000378:metabolism / Q000648:ultrastructure*; D029701:Saccharomyces cerevisiae Proteins; D008941:Spindle Apparatus
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; D009687:Nuclear Proteins; C073277:SPC110 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins