Effect of CCH1 and MID1 in calcium influx under alkaline pH and its regulation by Crz1p transcription factor in Candida albicans.
- Author:
Hui WANG
1
;
Ning XU
;
Laijun XING
;
Mingchun LI
;
Dongsheng WEI
Author Information
1. Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071, China.
- Publication Type:Journal Article
- MeSH:
Calcium Channels;
metabolism;
Candida albicans;
genetics;
metabolism;
physiology;
Fungal Proteins;
genetics;
physiology;
Gene Expression Regulation, Fungal;
Hydrogen-Ion Concentration;
Signal Transduction;
Stress, Physiological;
Transcription Factors;
metabolism;
physiology
- From:
Chinese Journal of Biotechnology
2011;27(6):917-925
- CountryChina
- Language:Chinese
-
Abstract:
In Candida albicans, adaptation to environmental pH is relevant to its pathogenicity. Calcium signaling pathway involves in many stress responses and often accompany with Ca2+ fluctuation. We constructed CCH1 and MID1 mutant strains and studied their effect on calcium influx and further investigated the regulation by Crz1p transcription factor. We used PCR-directed gene disruption to construct cch1delta/delta and mid1delta/delta null mutant. By using a flow cytometry-based method we monitored the free cytosolic Ca2+ levels under alkaline stress. Moreover, we constructed pPHO89-LacZ plasmids and by beta-Galactosidase assays, we analyzed the changes of LacZ activities after gene disruption. The results showed that alkaline stress induced calcium burst reduced obviously in cch1delta/delta and mid1delta/delta mutant strains, also for LacZ activities, and fully abolished in crz1delta/delta mutant strain. Finally, by realtime PCR, we confirmed the regulation role of Crz1p in CCH1 and MID1 genes but in a calcineurin independent way. Studies on the effect of calcium pathway on response to alkaline stress will provide an important theoretical basis for Candida albicans infection-oriented treatment and new drug targets.
