Direct measurement of tubulin and bulk message distributions on polysomes of growing, starved and deciliatedTetrahymenausing RNA gel blots of sucrose gradients containing acrylamide
| Autor: | Frank J. Calzone, Rosemary C. Callahan, Martin A. Gorovsky |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Acrylic Resins
Molecular Probe Techniques macromolecular substances Tubulin Polysome Gene expression Centrifugation Density Gradient Genetics Animals Centrifugation Cilia RNA Messenger Differential centrifugation Messenger RNA biology Tetrahymena Polynucleotide Adenylyltransferase RNA Blotting Northern Ethylenediamines biology.organism_classification Biochemistry Ammonium Sulfate Polyribosomes biology.protein Biophysics |
| Zdroj: | Nucleic Acids Research. 16:9597-9609 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/16.20.9597 |
| Popis: | A method was developed using sucrose gradients containing acrylamide which greatly simplifies the measurement of the polysomal distribution of messages. After centrifugation, the acrylamide was polymerized, forming a "polysome gel". RNA gel blots of polysome gels were used to determine the polysomal distributions of alpha-tubulin and total polyadenylated mRNA in growing, starved (nongrowing) and starved-deciliated Tetrahymena and the number of messages loaded onto polysomes was calculated. These measurements indicated that the translational efficiencies of alpha-tubulin mRNA and total polyadenylated mRNA are largely unaffected when the rates of tubulin and total protein synthesis vary dramatically. Thus, differential regulation of alpha-tubulin mRNA translation initiation does not contribute to the greater than 100-fold induction of tubulin synthesis observed during cilia regeneration and in growing cells. The major translation-level process regulating tubulin synthesis in Tetrahymena appears to be a change in message loading mediated by a non-specific message recruitment or unmasking factor. |
| Databáze: | OpenAIRE |
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