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Characterization of a putative spindle assembly checkpoint kinase Mps1, suggests its involvement in cell division, morphogenesis and oxidative stress tolerance in Candida albicans

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Title Characterization of a putative spindle assembly checkpoint kinase Mps1, suggests its involvement in cell division, morphogenesis and oxidative stress tolerance in Candida albicans
 
Creator Kamthan, Mohan
Nalla, Vijaya Kumar
Ruhela, Deepa
Kamthan, Ayushi
Maiti, Protiti
Datta, Asis
 
Subject Candida albicans
Oxidative Stress Tolerance
 
Description Accepted date: June 9, 2014
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 Efg1 and 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.
This work was supported by the Department of Biotechnology, the Ministry of Science and Technology, Government of India. The funders had no role
in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
 
Date 2015-12-17T04:45:22Z
2015-12-17T04:45:22Z
2014
 
Type Article
 
Identifier PLoS One, 9(7): e101517
1932-6203
http://172.16.0.77:8080/jspui/handle/123456789/418
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0101517
10.1371/journal.pone.0101517
 
Language en_US
 
Publisher PLOS