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MBC in Press, published online ahead of print March 5, 2004
Mol. Biol. Cell 10.1091/mbc.E03-10-0728

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Submitted on October 10, 2003
Revised on February 23, 2004
Accepted on February 23, 2004

Identification and characterization of two novel proteins affecting fission yeast {gamma}-tubulin complex function

Srinivas Venkatram1, Joseph J. Tasto1, Anna Feoktistova1, Jennifer L. Jennings2, Andrew J. Link2, and Kathleen L. Gould1*

1 HHMI, Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232
2 HHMI, Departments Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN 37232

* Corresponding author. E-mail address: kathy.gould{at}vanderbilt.edu.

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, mbo1{Delta} and gfh1{Delta} 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.




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