Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability

John S. Choy, Eileen O'Toole, Breanna M. Schuster, Matthew J. Crisp, Tatiana S. Karpova, James G. McNally, Mark Winey, Melissa K Gardner, Munira A. Basrai

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

How subunit dosage contributes to the assembly and function of multimeric complexes is an important question with implications in understanding biochemical, evolutionary, and disease mechanisms. Toward identifying pathways that are susceptible to decreased gene dosage, we performed a genome-wide screen for haploinsufficient (HI) genes that guard against genome instability in Saccharomyces cerevisiae. This led to the identification of all three genes (SPC97, SPC98, and TUB4) encoding the evolutionarily conserved γ-tubulin small complex (γ-TuSC), which nucleates microtubule assembly. We found that hemizygous γ-TuSC mutants exhibit higher rates of chromosome loss and increases in anaphase spindle length and elongation velocities. Fluorescence microscopy, fluorescence recovery after photobleaching, electron tomography, and model convolution simulation of spc98/+ mutants revealed improper regulation of interpolar (iMT) and kinetochore (kMT) microtubules in anaphase. The underlying cause is likely due to reduced levels of Tub4, as overexpression of TUB4 suppressed the spindle and chromosome segregation defects in spc98/+ mutants. We propose that γ-TuSC is crucial for balanced assembly between iMTs and kMTs for spindle organization and accurate chromosome segregation. Taken together, the results show how gene dosage studies provide critical insights into the assembly and function of multisubunit complexes that may not be revealed by using traditional studies with haploid gene deletion or conditional alleles.

Original languageEnglish (US)
Pages (from-to)2753-2763
Number of pages11
JournalMolecular biology of the cell
Volume24
Issue number17
DOIs
StatePublished - Sep 1 2013

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Haploinsufficiency
Chromosome Segregation
Anaphase
Gene Dosage
Genomic Instability
Microtubules
Electron Microscope Tomography
Genome
Fluorescence Recovery After Photobleaching
Kinetochores
Haploidy
Gene Deletion
Tubulin
Fluorescence Microscopy
Genes
Saccharomyces cerevisiae
Chromosomes
Alleles

Cite this

Choy, J. S., O'Toole, E., Schuster, B. M., Crisp, M. J., Karpova, T. S., McNally, J. G., ... Basrai, M. A. (2013). Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability. Molecular biology of the cell, 24(17), 2753-2763. https://doi.org/10.1091/mbc.E12-12-0902

Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability. / Choy, John S.; O'Toole, Eileen; Schuster, Breanna M.; Crisp, Matthew J.; Karpova, Tatiana S.; McNally, James G.; Winey, Mark; Gardner, Melissa K; Basrai, Munira A.

In: Molecular biology of the cell, Vol. 24, No. 17, 01.09.2013, p. 2753-2763.

Research output: Contribution to journalArticle

Choy, JS, O'Toole, E, Schuster, BM, Crisp, MJ, Karpova, TS, McNally, JG, Winey, M, Gardner, MK & Basrai, MA 2013, 'Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability', Molecular biology of the cell, vol. 24, no. 17, pp. 2753-2763. https://doi.org/10.1091/mbc.E12-12-0902
Choy, John S. ; O'Toole, Eileen ; Schuster, Breanna M. ; Crisp, Matthew J. ; Karpova, Tatiana S. ; McNally, James G. ; Winey, Mark ; Gardner, Melissa K ; Basrai, Munira A. / Genome-wide haploinsufficiency screen reveals a novel role for γ-TuSC in spindle organization and genome stability. In: Molecular biology of the cell. 2013 ; Vol. 24, No. 17. pp. 2753-2763.
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