Development of a yeast-based system to identify new hBRAFV600E functional interactors.

摘要: BRAFV600E is a mutant Ser–Thr protein kinase that plays crucial role in many types of cancer, including melanoma. Despite several aspects biology have been already elucidated, the proteins regulate its expression and activity remain largely unknown, hampering our capacity to control unrestrained effects. Here, we propose yeast Saccharomyces cerevisiae as model system can be used achieve better understanding regulation human BRAFV600E. By showing osmotic stress conditions hBRAFV600E rescue growth strains carrying double or triple deletion MAPKKK belonging HOG pathway, demonstrate this oncogenic active even if it does not an ortholog. Moreover, report that, ptp3∆ptc1∆ strain deleted genes encoding for two phosphatases responsible Hog1 de-phoshorylation, mimics toxicity observed presence constitutive activation. Finally, exploit such perform functional screening cDNA library, looking cDNAs able growth. In way, identify SMIM10, mitochondrial melanoma cells selectively downregulates RNA levels, by acting indirectly at post-transcriptional level. Upon SMIM10 overexpression, show disrupted structure/function undergo senescence. They also decreased ability proliferate form colonies, well increased sensitivity BRAF inhibitor vemurafenib. Interestingly, analysis TCGA samples indicates patients with higher levels prognosis. Therefore, these data suggest exerts oncosuppressive cells. Taken together, results unveil potential S. study hBRAFV600E, populate network interactors and, doing so, uncover new cancer-associated therapeutic potential.