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Cellular localization of RNA14p and RNA15p, two yeast proteins involved in mRNA stability
N Bonneaud1, L Minvielle-Sebastia, C Cullin
1Centre de Génétique Moléculaire du CNRS, Laboratoire Propre Associé à l'Université Pierre-et-Marie-Curie, Gif-sur-Yvette, France.
Abstract:
RNA14 and RNA15 were originally identified by temperature-sensitive mutations that cause a rapid decrease in poly(A)-tail length and overall mRNA levels at the restrictive temperature. We have raised antibodies to the RNA14 and RNA15 proteins, and used subcellular fractionation and immunofluorescence to localize these proteins within the yeast cell. RNA14p is a 73 kDa protein found in both the nucleus and the cytoplasm, whilst RNA15p is a 42 kDa protein detected only in the nucleus. The observation that both proteins are found in the nucleus is in agreement with previous genetic data which suggest an interaction between RNA14p and RNA15p. Also the joint nuclear localization is consistent with the biochemical data suggesting a role in polyadenylation. The detection of significant amounts of RNA14p in the cytoplasm opens the possibility of a second function for this protein, either in cytoplasmic regulation of mRNA deadenylation or, more interestingly, in mRNA stability.
Insights
Researchers localized RNA14 and RNA15 proteins in yeast cells. RNA14 is nuclear and cytoplasmic, while RNA15 is nuclear, suggesting roles in polyadenylation and potentially mRNA stability.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- RNA14 and RNA15 proteins were identified through temperature-sensitive mutations affecting poly(A)-tail length and mRNA levels.
- Previous genetic and biochemical data suggested an interaction between RNA14p and RNA15p, potentially involved in polyadenylation.
Purpose of the Study:
- To localize RNA14 and RNA15 proteins within the yeast cell using specific antibodies.
- To investigate the subcellular distribution of RNA14p and RNA15p to understand their cellular functions.
Main Methods:
- Antibodies were generated against RNA14 and RNA15 proteins.
- Subcellular fractionation and immunofluorescence techniques were employed for protein localization.
Main Results:
- RNA14 protein (73 kDa) was detected in both the nucleus and cytoplasm.
- RNA15 protein (42 kDa) was exclusively found in the nucleus.
- Nuclear localization of both proteins supports their interaction and role in polyadenylation.
Conclusions:
- The nuclear localization of RNA14p and RNA15p is consistent with their involvement in mRNA polyadenylation.
- The cytoplasmic presence of RNA14p suggests a potential additional role in cytoplasmic mRNA deadenylation or stability.