Mahon P, Beattie J, Glover L A, Hesketh J
Division of Biochemical Sciences, Rowett Research Institute, Aberdeen, Scotland, UK.
FEBS Lett. 1995 Oct 2;373(1):76-80. doi: 10.1016/0014-5793(95)01000-5.
The localisation of metallothionein isoform mRNAs in rat hepatoma (H4) cells was investigated using two approaches, namely Northern hybridisation of total RNA extracted from free, cytoskeletal-bound and membrane-bound polysomes isolated by a sequential detergent/salt extraction procedure and in situ hybridisation. The cytoskeletal-bound polysomes were enriched in metallothionein-I (MT-I) and c-myc mRNAs but showed a significantly lower enrichment in MT-II mRNA. These findings indicate that the MT-I mRNA is localised to the cytoskeleton during translation. In situ hybridisation using a biotin-labelled oligonucleotide probe revealed a predominantly perinuclear localisation for the MT-I mRNA.
采用两种方法研究了金属硫蛋白同工型mRNA在大鼠肝癌(H4)细胞中的定位,即对通过连续去污剂/盐提取程序分离的游离、细胞骨架结合和膜结合多聚核糖体中提取的总RNA进行Northern杂交以及原位杂交。细胞骨架结合的多聚核糖体富含金属硫蛋白-I(MT-I)和c-myc mRNA,但MT-II mRNA的富集程度明显较低。这些发现表明,MT-I mRNA在翻译过程中定位于细胞骨架。使用生物素标记的寡核苷酸探针进行的原位杂交显示,MT-I mRNA主要定位于核周。