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Yeast histone mRNA is polyadenylated
Abstract:
Histone mRNA from S. cerevisiae has been identified and partially purified. The RNA is quantitatively retained on oligo (dT) cellulose or poly(U) sepharose as assayed by in vitro translation or hybridization of radiolabelled cloned yeast histone sequences to RNA immobilized on DBM paper. Retention of yeast histone mRNA on either of these chromatographic systems is most likely the result of polyadenylation since, when primed with oligo (dT), the RNA is an extremely good template for reverse transcriptase, as determined by hybrid arrest translation or by hybridization to D. melanogaster histone DNA sequences.
Insights
Researchers identified and purified histone messenger RNA (mRNA) from yeast. This yeast histone mRNA exhibits polyadenylation, indicated by its retention on specific chromatography columns and its function as a template for reverse transcriptase.
Area of Science:
- Molecular Biology
- Biochemistry
- Yeast Genetics
Background:
- Histones are crucial proteins for DNA packaging in eukaryotes.
- Understanding histone gene expression is vital for cell biology.
- Messenger RNA (mRNA) carries genetic information for protein synthesis.
Purpose of the Study:
- To identify and partially purify histone mRNA from Saccharomyces cerevisiae (yeast).
- To investigate the characteristics of yeast histone mRNA, specifically its potential polyadenylation.
Main Methods:
- Isolation and partial purification of histone mRNA from S. cerevisiae.
- Chromatographic techniques using oligo (dT) cellulose and poly (U) sepharose for RNA retention assays.
- In vitro translation assays to assess mRNA activity.
- Hybridization techniques (DBM paper, D. melanogaster histone DNA) for RNA detection.
- Reverse transcriptase assays to evaluate template capabilities, primed with oligo (dT).
- Hybrid arrest translation experiments.
Main Results:
- Histone mRNA from S. cerevisiae was successfully identified and partially purified.
- The purified yeast histone mRNA was quantitatively retained on both oligo (dT) cellulose and poly (U) sepharose columns.
- The RNA demonstrated efficient template activity for reverse transcriptase when primed with oligo (dT).
- Hybridization studies confirmed the identity and characteristics of the retained mRNA.
Conclusions:
- Yeast histone mRNA possesses a polyadenylated tail, explaining its retention on affinity chromatography matrices.
- The polyadenylation of yeast histone mRNA suggests a regulatory mechanism potentially influencing its stability or translation.
- These findings contribute to the understanding of eukaryotic histone gene expression and mRNA processing.