Journal article
Complex Haploinsufficiency-Based Genetic Analysis of the NDR/Lats Kinase Cbk1 Provides Insight into Its Multiple Functions in Candida albicans
Genetics (Austin), Vol.203(3), pp.1217-1233
07/2016
DOI: 10.1534/genetics.116.188029
PMCID: PMC4937472
PMID: 27206715
Abstract
Although the analysis of genetic interactions and networks is a powerful approach to understanding biology, it has not been applied widely to the pathogenic yeast Candida albicans Here, we describe the use of both screening and directed genetic interaction studies based on complex haploinsufficiency to probe the function of the R: egulation of A: ce2 and M: orphogenesis (RAM) pathway in C. albicans A library of 5200 Tn7-mutagenized derivatives of a parental strain heterozygous at CBK1, the key kinase in the RAM pathway, was screened for alterations in serum-induced filamentation. Following confirmation of phenotypes and identification of insertion sites by sequencing, a set of 36 unique double heterozygous strains showing complex haploinsufficiency was obtained. In addition to a large set of genes regulated by the RAM transcription factor Ace2, genes related to cell wall biosynthesis, cell cycle, polarity, oxidative stress, and nitrogen utilization were identified. Follow-up analysis led to the first demonstration that the RAM pathway is required for oxidative stress tolerance in a manner related to the two-component-regulated kinase Chk1 and revealed a potential direct connection between the RAM pathway and the essential Mps1 spindle pole-related kinase. In addition, genetic interactions with CDC42-related genes MSB1, a putative scaffold protein, and RGD3, a putative Rho GTPase-activating protein (GAP) were identified. We also provide evidence that Rgd3 is a GAP for Cdc42 and show that its localization and phosphorylation are dependent on Cbk1.
Details
- Title: Subtitle
- Complex Haploinsufficiency-Based Genetic Analysis of the NDR/Lats Kinase Cbk1 Provides Insight into Its Multiple Functions in Candida albicans
- Creators
- Sarah Saputo - Department of Microbiology/Immunology University of Rochester School of Medicine and Dentistry, Rochester, New York 14642Kaitlyn L Norman - Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109Thomas Murante - Department of Pediatrics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642Brooke N Horton - Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109Jacinto De La Cruz Diaz - Department of Microbiology/Immunology University of Rochester School of Medicine and Dentistry, Rochester, New York 14642Louis DiDone - Department of Pediatrics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642Jennifer Colquhoun - Department of Microbiology/Immunology University of Rochester School of Medicine and Dentistry, Rochester, New York 14642Jeremy W Schroeder - Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109Lyle A Simmons - Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109Anuj Kumar - Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109Damian J Krysan - Department of Microbiology/Immunology University of Rochester School of Medicine and Dentistry, Rochester, New York 14642 Department of Pediatrics, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642 damian_krysan@urmc.rochester.edu
- Resource Type
- Journal article
- Publication Details
- Genetics (Austin), Vol.203(3), pp.1217-1233
- DOI
- 10.1534/genetics.116.188029
- PMID
- 27206715
- PMCID
- PMC4937472
- ISSN
- 0016-6731
- eISSN
- 1943-2631
- Grant note
- R25 GM064133 / NIGMS NIH HHS T90 DE021985 / NIDCR NIH HHS T32 GM007315 / NIGMS NIH HHS R01 AI098450 / NIAID NIH HHS T32 AI118689 / NIAID NIH HHS T32 GM007544 / NIGMS NIH HHS R01 GM107312 / NIGMS NIH HHS
- Language
- English
- Date published
- 07/2016
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Infectious Disease (Pediatrics)
- Record Identifier
- 9984093235602771
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