A protein transformation protocol for introducing yeast prion particles into yeast

Area of Science:

• Biochemistry
• Molecular Biology
• Yeast Genetics

Background:

• Established methods exist for nucleic acid transformation in organisms.
• Techniques for introducing proteins and protein aggregates into cells are less developed.

Purpose of the Study:

• To introduce a highly efficient protocol for introducing protein aggregates, such as prions, into yeast.
• To enable the study of prion inheritance by bridging in vitro and in vivo research.

Main Methods:

• Developed a protein transformation protocol for yeast.
• Infected yeast with recombinant Sup-NM amyloid fibers and in vivo Sup35p prions.
• Created distinct Sup-NM amyloid conformations for infection.

Main Results:

• Achieved high infectivity (approx. 100%) with Sup-NM fibers at high concentrations.
• Demonstrated that different Sup-NM amyloid conformations generate distinct yeast prion [PSI+] strains.
• Established a method adaptable to other prion proteins.

Conclusions:

• The protein transformation protocol is a significant advancement for studying protein aggregates and prions in yeast.
• This method facilitates the investigation of prion conformational diversity and inheritance mechanisms.
• The protocol bridges in vitro and in vivo prion research, aiding in understanding prion diseases.