The influence of mRNA localization on metallothionein-l protein distribution was studied by immunocytochemistry. We used Chinese hamster ovary cells that had been transfected with either a native metallothionein-l gene construct or metallothionein-l 5'-untranslated region and coding sequences linked to the 3'-untranslated region from glutathione peroxidase. The change in the 3'-untranslated region caused the delocalization of the mRNA with a loss of the perinuclear localization and association with the cytoskeleton. Clones were selected which expressed similar levels of metallothionein-l protein, as assessed by radioimmunoassay. The results showed that loss of metallothionein-l mRNA localization was associated with a loss of metallothionein-l protein localization, most notably with a lack of metallothionein-l protein in the nucleus of synchronized cells which were beginning to synthesize DNA This indicates that the association of metallothionein-l mRNA with the cytoskeleton around the nucleus is essential for efficient shuttling of the protein into the nucleus during the G(1) to S phase transition. This is the first demonstration of a physiological role for perinuclear mRNA localization and we propose that such localization may be important for a wide range of nuclear proteins, including those that shuttle between nucleus and cytoplasm in a cell cycle dependent manner.
|Number of pages||6|
|Journal||The Journal of Biological Chemistry|
|Publication status||Published - 1999|
- MESSENGER-RNA LOCALIZATION
- BOUND POLYSOMES
- 3'-UNTRANSLATED REGION
- RAT HEPATOCYTES