Zinc-dependent activation of the Pho8 alkaline phosphatase in Schizosaccharomyces pombe
| Autor: | Amanda J. Bird, Ya Mei Hu, Stevin Wilson, Hak Chung, Derek M. Boehm |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification 030102 biochemistry & molecular biology biology chemistry.chemical_element Cell Biology Zinc Vacuole Cycloheximide biology.organism_classification Biochemistry Transcriptome 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry Schizosaccharomyces pombe Metalloprotein Alkaline phosphatase Molecular Biology Cation diffusion facilitator |
| Zdroj: | Journal of Biological Chemistry. 294:12392-12404 |
| ISSN: | 0021-9258 |
| Popis: | Genome-wide analyses have revealed that during metal ion starvation, many cells undergo programmed changes in their transcriptome or proteome that lower the levels of abundant metalloproteins, conserving metal ions for more critical functions. Here we investigated how changes in cellular zinc status affect the expression and activity of the zinc-requiring Pho8 alkaline phosphatase from fission yeast (Schizosaccharomyces pombe). In S. pombe, Pho8 is a membrane-tethered and processed glycoprotein that resides in the vacuole. Using alkaline phosphatase activity assays along with various biochemical analyses, we found that Pho8 is active when zinc is plentiful and inactive when zinc is limited. Although Pho8 activity depended on zinc, we also found that higher levels of pho8 mRNAs and Pho8 protein accumulate in zinc-deficient cells. To gain a better understanding of the inverse relationship between pho8 mRNA levels and Pho8 activity, we examined the effects of zinc on the stability and processing of the Pho8 protein. We show that Pho8 is processed regardless of zinc status and that mature Pho8 accumulates under all conditions. We also noted that alkaline phosphatase activity is rapidly restored when zinc is resupplied to cells, even in the presence of the protein synthesis inhibitor cycloheximide. Our results suggest that S. pombe cells maintain inactive pools of Pho8 proteins under low-zinc conditions and that these pools facilitate rapid restoration of Pho8 activity when zinc ions become available. |
| Databáze: | OpenAIRE |
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