Identification and characterization of two novel proteins affecting fission yeast gamma-tubulin complex function.
The gamma-tubulin complex, via its ability to organize microtubules, is critical for accurate chromosome segregation and cytokinesis in the fission yeast, Schizosaccharomyces pombe. To better understand its roles, we have purified the S. pombe gamma-tubulin complex. Mass spectrometric analyses of the purified complex revealed known components and identified two novel proteins (i.e., Mbo1p and Gfh1p) with homology to gamma-tubulin-associated proteins from other organisms. We show that both Mbo1p and Gfh1p localize to microtubule organizing centers. Although cells deleted for either mbo1(+) or gfh1(+) are viable, they exhibit a number of defects associated with altered microtubule function such as defects in cell polarity, nuclear positioning, spindle orientation, and cleavage site specification. In addition, mbo1Delta and gfh1Delta cells exhibit defects in astral microtubule formation and anchoring, suggesting that these proteins have specific roles in astral microtubule function. This study expands the known roles of gamma-tubulin complex components in organizing different types of microtubule structures in S. pombe.
Department of Cell and Developmental Biology, Howard Hughes Medical Institute, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Srinivas
S
Venkatram
Joseph J
JJ
Tasto
Anna
A
Feoktistova
Jennifer L
JL
Jennings
Andrew J
AJ
Link
Kathleen L
KL
Gould
D001665:Binding Sites; D002148:Calmodulin-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D016764:Cell Polarity / Q000235:genetics; D017353:Gene Deletion; D049452:Green Fluorescent Proteins; D042581:Intranuclear Space / Q000648:ultrastructure; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D022101:Microtubule-Organizing Center / Q000378:metabolism / Q000502:physiology; D008870:Microtubules / Q000378:metabolism / Q000502:physiology; D008954:Models, Biological; D008969:Molecular Sequence Data; D025941:Protein Interaction Mapping; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D016415:Sequence Alignment; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism*
D002148:Calmodulin-Binding Proteins; C494594:Gfh1 protein, S pombe; C494593:Mbo1 protein, S pombe; D008869:Microtubule-Associated Proteins; D029702:Schizosaccharomyces pombe Proteins; D014404:Tubulin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins