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Quantitative Effects of Magnesium Chloride Stress on Aggregation of Sup35p in [psi-] Yeast Cells

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Abstract:

[PSI+] phenotype can be transiently induced when Magnesium chloride (MgCl2) was the selective pressure in SUP35 repeat-expansion mutant [psi-] yeast strains. We further investigated [PSI+] phenotype change under different MgCl2 conditions with native Sup35p and quantified the Sup35p status changes with fluorescence recovery after photobleaching (FRAP) and semi-denaturing detergent-agarose gel electrophoresis (SDD-AGE) analysis. It was found that the [PSI+] phenotype presented a dose-dependent relationship with the concentrations of MgCl2. Furthermore, Sup35p aggregated in MgCl2 treated cells but did not form large aggregates as it does in [PSI+] cells, and the size of Sup35p aggregates showed a time-dependent relationship with the MgCl2 application. The aggregation of Sup35p strictly depended on the presence of MgCl2 stress in our strains.





Keywords: Confocal Microscopy; FRAP assay; Green Fluorescent Protein; Immo-bilon polyvinylidene difluoride sheets; Immunoblotting; Live-Time Fluorescence Analysis; Magnesium chloride; NGMC diffusion; PSI+; SDD-AGE; SDS-PAGE; SDS-insoluble Sup35p; SDS-resistant Sup35p polymers; Saccharomyces cerevisiae; Sup35p; Sup35p in [psi-] Yeast Cells; Tris-borate-EDT; Western Blot Analysis; [PSI+]; adenine; anti-Sup35p antiserum; fluorescence recovery after photobleaching; guanidine curing; guanidine-hydrochloride; isogenic [Gpsi-] cells; nonsense suppression; prion; quantitative effects; semi-denaturing detergent-agarose gel electrophoresis; stress

Document Type: Research Article

Publication date: December 1, 2010

More about this publication?
  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.
ben/ppl/2010/00000017/00000012/art00005
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