Cell-cycle dependent phosphorylation of yeast pericentrin regulates γ-TuSC-mediated microtubule nucleation.
Budding yeast Spc110, a member of γ-tubulin complex receptor family (γ-TuCR), recruits γ-tubulin complexes to microtubule (MT) organizing centers (MTOCs). Biochemical studies suggest that Spc110 facilitates higher-order γ-tubulin complex assembly (Kollman et al., 2010). Nevertheless the molecular basis for this activity and the regulation are unclear. Here we show that Spc110 phosphorylated by Mps1 and Cdk1 activates γ-TuSC oligomerization and MT nucleation in a cell cycle dependent manner. Interaction between the N-terminus of the γ-TuSC subunit Spc98 and Spc110 is important for this activity. Besides the conserved CM1 motif in γ-TuCRs (Sawin et al., 2004), a second motif that we named Spc110/Pcp1 motif (SPM) is also important for MT nucleation. The activating Mps1 and Cdk1 sites lie between SPM and CM1 motifs. Most organisms have both SPM-CM1 (Spc110/Pcp1/PCNT) and CM1-only (Spc72/Mto1/Cnn/CDK5RAP2/myomegalin) types of γ-TuCRs. The two types of γ-TuCRs contain distinct but conserved C-terminal MTOC targeting domains.DOI: http://dx.doi.org/10.7554/eLife.02208.001.
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany The Hartmut Hoffmann-Berling International Graduate School, University of Heidelberg, Heidelberg, Germany.
Tien-Chen
TC
Lin
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany.
Annett
A
Neuner
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany.
Yvonne T
YT
Schlosser
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany.
Annette N D
AN
Scharf
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany.
Lisa
L
Weber
Zentrum für Molekulare Biologie (ZMBH), Universität Heidelberg, Heidelberg, Germany e.schiebel@zmbh.uni-heidelberg.de.
Elmar
E
Schiebel
D000595:Amino Acid Sequence; D000941:Antigens / Q000502:physiology*; D001704:Biopolymers / Q000378:metabolism; D002453:Cell Cycle; D008870:Microtubules / Q000502:physiology*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017386:Sequence Homology, Amino Acid; D014404:Tubulin / Q000737:chemistry / Q000502:physiology*
D000941:Antigens; D001704:Biopolymers; D014404:Tubulin; C086178:pericentrin
gamma-tubulin complex; microtubule nucleation; mitosis; pericentrin; phosphorylation; spindle pole body