Abstract
Cobalt is an essential micronutrient but is toxic when present in excess. To study cobalt homeostasis we performed a genome-wide screen for deletion strains that show sensitivity or resistance to CoCl2. Among 54 cobalt-sensitive strains, 18 are supersensitive strains, which are involved in histidine biosynthetic process, ubiquitination, mitochondria function, membrane trafficking, transporter and a variety of other known functions or still unknown functions. Furthermore, we identified 56 cobalt-resistant deletion strains, which are mainly involved in mitochondria function, signal transduction, ubiquitination, and gene expression and chromatin remodeling. Notably, deletion of the zhf1 + gene, encoding a zinc ion transporter, confers supersensitivity to cobalt and overexpression of the zhf1 + gene confers marked tolerance to cobalt, indicating that Zhf1 play key roles in cobalt detoxification. Interestingly, all the histidine-auxotrophic mutants displayed cobalt sensitivity and deletion of cationic amino acid transporter Cat1, which was shown to be involved in histidine uptake, suppressed the CoCl2-sensitive growth defect of the his2 mutants, suggesting that CoCl2 may be transported into the cell together with histidine via histidine transporters including Cat1. In addition, we obtained results suggesting that the E2 ubiquitin conjugating enzyme Rhp6 and Sty1 stress MAP kinase pathway are involved in the regulation of cobalt homeostasis. Altogether, our genome-wide study demonstrates for the first time the mechanisms of cobalt homeostasis, particularly its uptake and detoxification in fission yeast.
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Abbreviations
- CRE:
-
cAMP-response element
- EMM:
-
Edinburgh minimal medium
- MAPK:
-
Mitogen-activated protein kinase
- ORF:
-
Open reading frame
- RLU:
-
Relative light units
- YES:
-
Yeast extract with supplements
- YPD:
-
Yeast extract–peptone–dextrose
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Acknowledgments
We thank Susie O. Sio for critical reading of the manuscript, and Kayo Osaka for her contribution to the preliminary stages of this study. Financial support for this study was provided by Grant-in-Aid for Scientific Research, on Scientific Research (B) to Takayoshi Kuno (No. 22390048) and on Grant-in-Aid for Young Scientists (B) to Yan Ma (No. 21790241), from Japan Society for the Promotion of Science (http://www.jsps.go.jp/english/e-grants/grants.html).
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Communicated by T. Ito.
S. Ryuko and Y. Ma contributed equally to this work.
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Ryuko, S., Ma, Y., Ma, N. et al. Genome-wide screen reveals novel mechanisms for regulating cobalt uptake and detoxification in fission yeast. Mol Genet Genomics 287, 651–662 (2012). https://doi.org/10.1007/s00438-012-0705-9
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DOI: https://doi.org/10.1007/s00438-012-0705-9