Direct Observation of Yeast Prion Dynamics in Single-Living Cells

摘要: Since prions are transmissible, they inherently replicate themselves in order to propagate the transmissible entities, and then transmit daughter cells. To understand dynamics of yeast prion aggregates living cells, we directly monitored fate using an on-chip single-cell cultivation system conjunction with fluorescence correlation spectroscopy (FCS), a technique that allows determination diffusion times fluorescent molecules even Single-cell imaging revealed visible foci Sup35 fused GFP dispersed throughout cytoplasm during cell growth, but retain phenotype. Fluorescent spectroscopy, which showed [PSI + ] irrespective presence foci, contain diffuse oligomers, transmitted their We concluded these oligomers critical for transmission. FCS is ensemble method calculating properties molecules, it cannot be used track behavior individual molecules. overcome this limitation, extending our study dynamic single proteins by quantum dot (Q-dot) technology. Q-dots semiconductor nanocrystals have several advantages over conventional organic dyes. chemically labeled recombinant Q-dots, incorporated Q-dot labelled (Sup35-Qdot) into succeeded tracking trajectories Sup35-Qdots analyzed properties.