Ppc89 links multiple proteins, including the septation initiation network, to the core of the fission yeast spindle-pole body.
The spindle-pole body (SPB), the yeast analog of the centrosome, serves as the major microtubule (MT) organizing center in the yeast cell. In addition to this central function, the SPB organizes and concentrates proteins required for proper coordination between the nuclear-division cycle and cytokinesis. For example, the Schizosaccharomyces pombe septation-initiation network (SIN), which is responsible for initiating actomyosin ring constriction and septation, is assembled at the SPB through its two scaffolding components, Sid4 and Cdc11. In an effort to identify novel SIN interactors, we purified Cdc11 and identified by mass spectrometry a previously uncharacterized protein associated with it, Ppc89. Ppc89 localizes constitutively to the SPB and interacts directly with Sid4. A fusion between the N-terminal 300 amino acids of Sid4 and a SPB targeting domain of Ppc89 supplies the essential function of Sid4 in anchoring the SIN. ppc89Delta cells are inviable and exhibit defects in SPB integrity, and hence in spindle formation, chromosome segregation, and SIN localization. Ppc89 overproduction is lethal, resulting primarily in a G2 arrest accompanied by massive enlargement of the SPB and increased SPB MT nucleation. These results suggest a fundamental role for Ppc89 in organization of the S. pombe SPB.
Rosenberg
Joshua A
JA
Howard Hughes Medical Institute and Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Tomlin
Gregory C
GC
McDonald
W Hayes
WH
Snydsman
Brian E
BE
Muller
Eric G
EG
Yates
John R
JR
Gould
Kathleen L
KL
D031541:Fluorescence Resonance Energy Transfer; D017353:Gene Deletion; D015870:Gene Expression; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000378:metabolism*; D008941:Spindle Apparatus / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure
D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D029702:Schizosaccharomyces pombe Proteins; C409024:Sid4 protein, S pombe