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[Characterization of messenger ribonucleoproteins from yeasts (author's transl)]
Zeitschrift Fur Naturforschung. Section C, Biosciences
|July 1, 1975
Abstract:
A method to produce in 15 min stable sphaeroplasts from yeast of the early log-phase is described. This method was used to isolate polysomes, after specific radioactive labelling of mRNA. The mRNP *-particles, isolated from polysomes, have sedimentation-coefficients from 10 to 80S and a density of 1.4 g/cm3, the polysomes a density of 1.58 g/cm3 and the ribosomes a density of 1.6. g/cm3.
Insights
Researchers developed a rapid 15-minute method to create stable yeast spheroplasts. This technique enables efficient isolation of polysomes and messenger ribonucleoprotein (mRNP) particles for molecular studies.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Yeast spheroplasts are crucial for studying cellular processes like mRNA metabolism.
- Previous methods for yeast spheroplast preparation were time-consuming.
- Efficient isolation of polysomes and associated mRNP particles is key to understanding gene expression.
Purpose of the Study:
- To establish a rapid and reliable method for generating stable yeast spheroplasts.
- To utilize these spheroplasts for the isolation of polysomes and mRNP particles.
- To characterize the biophysical properties of isolated polysomes and mRNP particles.
Main Methods:
- Development of a 15-minute protocol for producing stable spheroplasts from early log-phase yeast.
- Application of the spheroplast method for isolating polysomes following radioactive mRNA labeling.
- Isolation and characterization of messenger ribonucleoprotein (mRNP) particles from polysomes using ultracentrifugation.
Main Results:
- Stable yeast spheroplasts were successfully produced within 15 minutes.
- The method allowed for the isolation of polysomes from specifically labeled mRNA.
- Isolated mRNP particles exhibited sedimentation coefficients of 10–80S and a density of 1.4 g/cm³.
- Polysomes and ribosomes showed densities of 1.58 g/cm³ and 1.6 g/cm³, respectively.
Conclusions:
- The described method provides a fast and effective way to obtain stable yeast spheroplasts.
- This technique facilitates the study of mRNA metabolism and protein synthesis in yeast.
- Characterization of mRNP particles and polysomes provides insights into their molecular composition and dynamics.