Modular control of cross-oligomerization: analysis of superstabilized Hsp90 homodimers in vivo.

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摘要：

Homo-oligomeric proteins fulfill numerous functions in all cells. The ability to co-express subunits of these proteins that preferentially self-assemble without cross-oligomerizing provides for controlled experiments to analyze the function of mutant homo-oligomers in vivo. is a dimeric chaperone involved in the maturation of many kinases and steroid hormone receptors. We observed that co-expression of different subunits in caused unpredictable synthetic growth defects due to cross-dimerization. We engineered superstabilized dimers that resisted cross-dimerization with endogenous and alleviated the synthetic growth defect. Superstabilized dimers supported robust growth of , indicating that dissociation of dimers could be hindered without compromising essential function. We utilized superstabilized dimers to analyze the activity of ATPase mutant homodimers in a temperature-sensitive background where elevated temperature inactivated all other species. We found that and hydrolysis by are both required for the efficient maturation of and , confirming the critical role of in the maturation of steroid hormone receptors and kinases in vivo.

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Saccharomyces cerevisiae Oncogene Protein pp60(v-src Receptors, Glucocorticoid Temperature Protein Structure, Quaternary Protein Structure, Secondary Adenosine Triphosphatases Mutant Proteins Transcriptional Activation Protein Multimerization