Investigation On The Effects Of Ribosomal Protein Mutations On Stress Resistance And Lifespan Extension In Yeast

Ribosome is one of the most important organelles in cellsand and it‘s the major site of protein synthesis.When some of ribosome protein expression of an exception occurs,such as loss or excessive expression,or change its structure,these will affect the assembly of the ribosome,resulting in abnormal protein synthesis,which affect cell function.At the early stages of our work,we test the growth conditions of FY251,?36B,2A,2B,?36A,?11A,?11B with Spotting assay.AS we can see knockingout some ribosomal protein such as RPL36 B RPL11A had significant effect on the growth of bacteria,and knocking out their homologous genes RPL36 A RPL11B effects on bacteria are not obvious,and the function of homologous RNA protein is not exactly the same.Then we test the growth curve of these yeast for a long time,and find the growth of ?36B is not that bad.At first,there is a severe growth defect of?36B but the growth of ?36B is similar to FY251 after 18 h and much more betterthan FY251 after 36 h.Why ?36B grown much better with the consumption ofnutrients and the steady accumulation of metabolites?IS there a similar effect when knockdown the large subnit of ribosome protein?we test growth curve of knockdown of RPL7 A,RPL7B,RPL11 A,RPL11B as well,and find no that growth rule like?36B.To find what makes ?36B grown much better after 36 h,we test the growth ofsome yeast which RPL36 B knockeddown and RPL36 A have changed.Such as replece RPL36 A promoter with ADH1 promoter,knockdown the introns of RPL36 A,or add protein tag flag to the end of C-terminal gene RPL36 A.And there is no similar growth effect with ?36B.We also examined the expression of ribosome protein,accumulation of reactive oxygen species(ROS),LacZ activity,nutrient limitation and protein aggregate to find the reason for growth of ?36B.There is a large accumulation of unfolded protein ormisfold protein at the earlier stage of the growth of ?36B,but less after 36 h.So weconsider the aggregation of protein is one of the main factors for the lifespan of yeast.With the optimization of the experimental tools and deepening of the experiments,it is hopeful there is more to be learned about the special growth of ?36B.