Palmitoylation of the Cysteine Residue in the DHHC Motif of a Palmitoyl Transferase Mediates Ca2+ Homeostasis in Aspergillus
| Autor: | Lina Gao, Shizhu Zhang, Congcong Sun, Nick D. Read, Yuanwei Zhang, Ling Lu, Qing-qing Zheng, Alberto Muñoz, Jinxing Song |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Hyphal growth Aspergillus Nidulans Cancer Research Antifungal Agents Physiology ATPase Cell Membranes Amino Acid Motifs Yeast and Fungal Models Biochemistry Cell Signaling Medicine and Health Sciences Homeostasis Post-Translational Modification Amino Acids Genetics (clinical) Calcium signaling Adenosine Triphosphatases biology Organic Compounds Tunicamycin Spores Fungal Endoplasmic Reticulum Stress Cell biology Chemistry Aspergillus Physical Sciences Cellular Structures and Organelles Itraconazole Palmitoylation Research Article Signal Transduction Proton ATPase lcsh:QH426-470 Bone and Mineral Metabolism Lipoylation 030106 microbiology Hyphae chemistry.chemical_element Calcium Research and Analysis Methods 03 medical and health sciences Model Organisms Genetics Extracellular Sulfur Containing Amino Acids Calcium Signaling Cysteine Molecular Biology Ecology Evolution Behavior and Systematics Ion Transport Organic Chemistry Organisms Fungi Chemical Compounds Biology and Life Sciences Proteins Membrane Proteins Cell Biology Molds (Fungi) lcsh:Genetics Metabolism Membrane protein chemistry biology.protein Physiological Processes Acyltransferases |
| Zdroj: | PLoS Genetics PLoS Genetics, Vol 12, Iss 4, p e1005977 (2016) |
| ISSN: | 1553-7404 1553-7390 |
| Popis: | Finely tuned changes in cytosolic free calcium ([Ca2+]c) mediate numerous intracellular functions resulting in the activation or inactivation of a series of target proteins. Palmitoylation is a reversible post-translational modification involved in membrane protein trafficking between membranes and in their functional modulation. However, studies on the relationship between palmitoylation and calcium signaling have been limited. Here, we demonstrate that the yeast palmitoyl transferase ScAkr1p homolog, AkrA in Aspergillus nidulans, regulates [Ca2+]c homeostasis. Deletion of akrA showed marked defects in hyphal growth and conidiation under low calcium conditions which were similar to the effects of deleting components of the high-affinity calcium uptake system (HACS). The [Ca2+]c dynamics in living cells expressing the calcium reporter aequorin in different akrA mutant backgrounds were defective in their [Ca2+]c responses to high extracellular Ca2+ stress or drugs that cause ER or plasma membrane stress. All of these effects on the [Ca2+]c responses mediated by AkrA were closely associated with the cysteine residue of the AkrA DHHC motif, which is required for palmitoylation by AkrA. Using the acyl-biotin exchange chemistry assay combined with proteomic mass spectrometry, we identified protein substrates palmitoylated by AkrA including two new putative P-type ATPases (Pmc1 and Spf1 homologs), a putative proton V-type proton ATPase (Vma5 homolog) and three putative proteins in A. nidulans, the transcripts of which have previously been shown to be induced by extracellular calcium stress in a CrzA-dependent manner. Thus, our findings provide strong evidence that the AkrA protein regulates [Ca2+]c homeostasis by palmitoylating these protein candidates and give new insights the role of palmitoylation in the regulation of calcium-mediated responses to extracellular, ER or plasma membrane stress. Author Summary Palmitoylation is a reversible post-translational modification catalyzed by palmitoyl acyltransferases (PATs) and proteins that undergo this modification are involved in numerous intracellular functions. Yeast Akr1p was the first characterized PAT whilst HIP14, an Akr1p homolog in human, is one of the most highly conserved of 23 human PATs that catalyze the addition of palmitate to the Huntington protein which is of major importance in Huntington’s disease. Calcium serves numerous signaling and structural functions in all eukaryotes. However, studies on the relationship between calcium signaling and palmitoylation are lacking. In this study, we demonstrate that the palmitoyl transferase Akr1 homolog in the filamentous fungus Aspergillus nidulans, similar to the high-affinity calcium uptake system (HACS), is required for normal growth and sporulation in the presence of low extracellular calcium. We find that AkrA dysfunction decreases the transient increase in cytosolic free calcium induced by a high extracellular calcium stress, tunicamycin (which induces endoplasmic reticulum stress) or the antifungal agent itraconazole (which induces plasma membrane stress). The influence of AkrA on all of these processes involves its DHHC motif, which is required for palmitoylation of various proteins associated with many processes including calcium signaling and membrane trafficking. Our findings provide evidence for a crucial link between calcium signaling and palmitoylation, suggesting a possible role in the mechanistic basis of human PAT-related diseases. These results also indicate that regulators of posttranslational modification may provide promising antifungal targets for new therapies. |
| Databáze: | OpenAIRE |
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