Genetics, Vol 137, 407-422, Copyright © 1994

INVESTIGATIONS

Genetic Interactions Between CDC31 and KAR1, Two Genes Required for Duplication of the Microtubule Organizing Center in Saccharomyces cerevisiae

E. A. Vallen, W. Ho, M. Winey and M. D. Rose
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544-1014

KAR1 encodes an essential component of the yeast spindle pole body (SPB) that is required for karyogamy and SPB duplication. A temperature-sensitive mutation, kar1-{Delta}17, mapped to a region required for SPB duplication and for localization to the SPB. To identify interacting SPB proteins, we isolated 13 dominant mutations and 13 high copy number plasmids that suppressed the temperature sensitivity of kar1-{Delta}17. Eleven extragenic suppressor mutations mapped to two linkage groups, DSK1 and DSK2. The extragenic suppressors were specific for SPB duplication and did not suppress karyogamy-defective alleles. The major class, DSK1, consisted of mutations in CDC31. CDC31 is required for SPB duplication and encodes a calmodulin-like protein that is most closely related to caltractin/centrin, a protein associated with the Chlamydomonas basal body. The high copy number suppressor plasmids contained the wild-type CDC31 gene. One CDC31 suppressor allele conferred a temperature-sensitive defect in SPB duplication, which was counter-suppressed by recessive mutations in KAR1. In spite of the evidence for a direct interaction, the strongest CDC31 alleles, as well as both DSK2 alleles, suppressed a complete deletion of KAR1. However, the CDC31 alleles also made the cell supersensitive to KAR1 gene dosage, arguing against a simple bypass mechanism of suppression. We propose a model in which Kar1p helps localize Cdc31p to the SPB and that Cdc31p then initiates SPB duplication via interaction with a downstream effector.

This article has been cited by other articles:

				L. Chen and K. Madura Centrin/Cdc31 Is a Novel Regulator of Protein Degradation Mol. Cell. Biol., March 1, 2008; 28(5): 1829 - 1840. [Abstract] [Full Text] [PDF]

				S. L. Jaspersen, A. E. Martin, G. Glazko, T. H. Giddings Jr., G. Morgan, A. Mushegian, and M. Winey The Sad1-UNC-84 homology domain in Mps3 interacts with Mps2 to connect the spindle pole body with the nuclear envelope J. Cell Biol., August 28, 2006; 174(5): 665 - 675. [Abstract] [Full Text] [PDF]

				S. Li, A. M. Sandercock, P. Conduit, C. V. Robinson, R. L. Williams, and J. V. Kilmartin Structural role of Sfi1p-centrin filaments in budding yeast spindle pole body duplication J. Cell Biol., June 19, 2006; 173(6): 867 - 877. [Abstract] [Full Text] [PDF]

				A. Selvapandiyan, A. Debrabant, R. Duncan, J. Muller, P. Salotra, G. Sreenivas, J. L. Salisbury, and H. L. Nakhasi Centrin Gene Disruption Impairs Stage-specific Basal Body Duplication and Cell Cycle Progression in Leishmania J. Biol. Chem., June 11, 2004; 279(24): 25703 - 25710. [Abstract] [Full Text] [PDF]

				A. Popescu, S. Miron, Y. Blouquit, P. Duchambon, P. Christova, and C. T. Craescu Xeroderma Pigmentosum Group C Protein Possesses a High Affinity Binding Site to Human Centrin 2 and Calmodulin J. Biol. Chem., October 10, 2003; 278(41): 40252 - 40261. [Abstract] [Full Text] [PDF]

				J. V. Kilmartin Sfi1p has conserved centrin-binding sites and an essential function in budding yeast spindle pole body duplication J. Cell Biol., September 29, 2003; 162(7): 1211 - 1221. [Abstract] [Full Text] [PDF]

				O. Gavet, C. Alvarez, P. Gaspar, and M. Bornens Centrin4p, a Novel Mammalian Centrin Specifically Expressed in Ciliated Cells Mol. Biol. Cell, May 1, 2003; 14(5): 1818 - 1834. [Abstract] [Full Text] [PDF]

				K. Lee, L. Neigeborn, and R. J. Kaufman The Unfolded Protein Response Is Required for Haploid Tolerance in Yeast J. Biol. Chem., March 28, 2003; 278(14): 11818 - 11827. [Abstract] [Full Text] [PDF]

				S. L. Jaspersen, T. H. Giddings Jr., and M. Winey Mps3p is a novel component of the yeast spindle pole body that interacts with the yeast centrin homologue Cdc31p J. Cell Biol., December 23, 2002; 159(6): 945 - 956. [Abstract] [Full Text] [PDF]

				A. Selvapandiyan, R. Duncan, A. Debrabant, S. Bertholet, G. Sreenivas, N. S. Negi, P. Salotra, and H. L. Nakhasi Expression of a Mutant Form of Leishmania donovani Centrin Reduces the Growth of the Parasite J. Biol. Chem., November 9, 2001; 276(46): 43253 - 43261. [Abstract] [Full Text] [PDF]

				I. Ivanovska and M. D. Rose Fine Structure Analysis of the Yeast Centrin, Cdc31p, Identifies Residues Specific for Cell Morphology and Spindle Pole Body Duplication Genetics, February 1, 2001; 157(2): 503 - 518. [Abstract] [Full Text]

				W. Khalfan, I. Ivanovska, and M. D. Rose Functional Interaction Between the PKC1 Pathway and CDC31 Network of SPB Duplication Genes Genetics, August 1, 2000; 155(4): 1543 - 1559. [Abstract] [Full Text]

				S. Middendorp, T. Kuntziger, Y. Abraham, S. Holmes, N. Bordes, M. Paintrand, A. Paoletti, and M. Bornens A Role for Centrin 3 in Centrosome Reproduction J. Cell Biol., February 7, 2000; 148(3): 405 - 416. [Abstract] [Full Text] [PDF]

				T. M. Sandrock, S. M. Brower, K. A. Toenjes, and A. E. M. Adams Suppressor Analysis of Fimbrin (Sac6p) Overexpression in Yeast Genetics, April 1, 1999; 151(4): 1287 - 1297. [Abstract] [Full Text]

				D. S. Sullivan, S. Biggins, and M. D. Rose The Yeast Centrin, Cdc31p, and the Interacting Protein Kinase, Kic1p, Are Required for Cell Integrity J. Cell Biol., November 2, 1998; 143(3): 751 - 765. [Abstract] [Full Text] [PDF]

				J. Petersen, O. Nielsen, R. Egel, and I. M. Hagan FH3, A Domain Found in Formins, Targets the Fission Yeast Formin Fus1 to the Projection Tip During Conjugation J. Cell Biol., June 1, 1998; 141(5): 1217 - 1228. [Abstract] [Full Text] [PDF]

				R. D.C. Saunders, M. d. C. Avides, T. Howard, C. Gonzalez, and D. M. Glover The Drosophila Gene abnormal spindle Encodes a Novel Microtubule-associated Protein That Associates with the Polar Regions of the Mitotic Spindle J. Cell Biol., May 19, 1997; 137(4): 881 - 890. [Abstract] [Full Text] [PDF]

				B. M. Geier, H. Wiech, and E. Schiebel Binding of Centrins and Yeast Calmodulin to Synthetic Peptides Corresponding to Binding Sites in the Spindle Pole Body Components Kar1p and Spc110p J. Biol. Chem., November 8, 1996; 271(45): 28366 - 28374. [Abstract] [Full Text] [PDF]

				H. Wiech, B. M. Geier, T. Paschke, A. Spang, K. Grein, J. Steinkotter, M. Melkonian, and E. Schiebel Characterization of Green Alga, Yeast, and Human Centrins. SPECIFIC SUBDOMAIN FEATURES DETERMINE FUNCTIONAL DIVERSITY J. Biol. Chem., September 13, 1996; 271(37): 22453 - 22461. [Abstract] [Full Text] [PDF]

				M. Araki, C. Masutani, M. Takemura, A. Uchida, K. Sugasawa, J. Kondoh, Y. Ohkuma, and F. Hanaoka Centrosome Protein Centrin 2/Caltractin 1 Is Part of the Xeroderma Pigmentosum Group C Complex That Initiates Global Genome Nucleotide Excision Repair J. Biol. Chem., May 25, 2001; 276(22): 18665 - 18672. [Abstract] [Full Text] [PDF]