Mutation of the Ser18 phosphorylation site on the sole Saccharomyces cerevisiae UCS protein, She4, can compromise high-temperature survival
| Autor: | Susana Gomez-Escalante, Stefan H. Millson, Peter W. Piper |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Saccharomyces cerevisiae Proteins Amino Acid Motifs Mutant Saccharomyces cerevisiae Hsp90 Biochemistry Myosin Type I 03 medical and health sciences She4 Two-Hybrid System Techniques Myosin UCS proteins HSP90 Heat-Shock Proteins Cytoskeleton Original Paper UNC45 030102 biochemistry & molecular biology biology C130 Cell Biology Temperature Cell Biology biology.organism_classification Endocytosis Yeast C510 Applied Microbiology Cytoskeletal Proteins Phenotype 030104 developmental biology Microscopy Fluorescence Temperature stress Mutagenesis Site-Directed biology.protein Phosphorylation Sequence motif |
| Zdroj: | Cell Stress & Chaperones |
| ISSN: | 1466-1268 1355-8145 |
| Popis: | Folding of the myosin head often requires the joint actions of Hsp90 and a dedicated UNC45, Cro1, She4 (UCS) domain-containing cochaperone protein. Relatively weak sequence conservation exists between the single UCS protein of simple eukaryotes (She4 in budding yeast) and the two UCS proteins of higher organisms (the general cell and smooth muscle UNC45s; UNC45-GC and UNC45-SM respectively). In vertebrates, UNC45-GC facilitates cytoskeletal function whereas the 55% identical UNC45-SM assists in the assembly of the contractile apparatus of cardiac and skeletal muscles. UNC45-SM, unlike UNC45-GC, shares with yeast She4 an IDSL sequence motif known to be a site of in vivo serine phosphorylation in yeast. Investigating this further, we found that both a non-phosphorylatable (S18A) and a phosphomimetic (S18E) mutant form of She4 could rescue the type 1 myosin localisation and endocytosis defects of the yeast she4Δ mutant at 39 °C. Nevertheless, at higher temperature (45 °C), only She4 (S18A), not She4(S18E), could substantially rescue the cell lysis defect of she4Δ mutant cells. In the yeast two-hybrid system, the non-phosphorylatable S18A and S251A mutant forms of She4 and UNC45-SM still displayed the stress-enhanced in vivo interaction with Hsp90 seen with the wild-type She4 and UNC45-SM. Such high-temperature enforcement to interaction was though lost with the phosphomimetic mutant forms (She4(S18E) and UNC45-SM (S251E)), an indication that phosphorylation might suppress these increases in She4/Hsp90 and UNC45-SM/Hsp90 interaction with stress. |
| Databáze: | OpenAIRE |
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