Hovland R, Campbell G, Pryme I, Hesketh J
Department of Biochemistry and Molecular Biology, University of Bergen, Norway.
Biochem J. 1995 Aug 15;310 ( Pt 1)(Pt 1):193-6. doi: 10.1042/bj3100193.
Cytosolic, cytoskeleton and membrane fractions were extracted from HepG2 cells by a sequential detergent/salt extraction procedure. The cytosolic fraction contained 93% of the lactic dehydrogenase activity while the cytoskeleton fraction was enriched in actin and vimentin. The distribution of mRNAs for c-myc, glucose transporter 1, ribosomal proteins L4 and S6 and cyclin A were investigated by Northern hybridization of total RNA extracted from polysomes isolated from cytosolic, cytoskeleton and membrane fractions. The membrane-bound polysomes were enriched in the glucose transporter 1 mRNA and the cytoskeleton-bound polysomes were enriched in the mRNAs for the two ribosomal proteins, c-myc and cyclin A. The results suggest that the mRNAs for nuclear proteins are one class of mRNAs which are translated on polysomes associated with the cytoskeleton; this may be related to the requirement to transport the newly synthesized protein to the nucleus.
采用连续的去污剂/盐提取法从HepG2细胞中提取胞质、细胞骨架和膜组分。胞质组分含有93%的乳酸脱氢酶活性,而细胞骨架组分富含肌动蛋白和波形蛋白。通过对从胞质、细胞骨架和膜组分分离的多聚核糖体中提取的总RNA进行Northern杂交,研究了c-myc、葡萄糖转运蛋白1、核糖体蛋白L4和S6以及细胞周期蛋白A的mRNA分布。膜结合多聚核糖体富含葡萄糖转运蛋白1 mRNA,细胞骨架结合多聚核糖体富含两种核糖体蛋白、c-myc和细胞周期蛋白A的mRNA。结果表明,核蛋白的mRNA是一类在与细胞骨架相关的多聚核糖体上翻译的mRNA;这可能与将新合成的蛋白质转运到细胞核的需求有关。