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In  Saccharomyces cerevisiae MPS1  is one of the major protein kinase that governs the spindle checkpoint pathway. The  S. cerevisiae  structural homolog of opportunistic pathogen  Candida albicans CaMPS1 , is indispensable for the cell viability. The essentiality of Mps1 was confirmed by Homozygote Trisome test. To determine its biological function in this pathogen conditional mutant was generated through regulatable  MET3  promoter. Examination of heterozygous and conditional (+Met/Cys)  mps1  mutants revealed a mitosis specific arrest phenotype, where mutants showed large buds with undivided nuclei. Flowcytometry analysis revealed abnormal ploidy levels in  mps1 mutant. In presence of anti-microtubule drug Nocodazole,  mps1  mutant showed a dramatic loss of viability suggesting a role of Mps1 in Spindle Assembly Checkpoint (SAC) activation. These mutants were also defective in microtubule organization. Moreover, heterozygous mutant showed defective in-vitro yeast to hyphae morphological transition. Growth defect in heterozygous mutant suggest haploinsufficiency of this gene. qRT PCR analysis showed around 3 fold upregulation of  MPS1  in presence of serum. This expression of  MPS1  is dependent on Efg1and is independent of other hyphal regulators like Ras1 and Tpk2. Furthermore,  mps1  mutants were also sensitive to oxidative stress. Heterozygous  mps1  mutant did not undergo morphological transition and showed 5-Fold reduction in colony forming units in response to macrophage. Thus, the vital checkpoint kinase, Mps1 besides cell division also has a role in morphogenesis and oxidative stress tolerance, in this pathogenic fungus.

plos one

Characterization of a Putative Spindle Assembly Checkpoint Kinase Mps1, Suggests Its Involvement in Cell Division, Morphogenesis and Oxidative Stress Tolerance in Candida albicans