Müller W E, Agutter P S, Prochnow D J, Fasold H, Sève A P, Tsiapalis C M, Schröder H C
Institut für Physiologische Chemie, Abteilung für Angewandte Molekularbiologie, Mainz, Germany.
Z Gerontol. 1993 Jul-Aug;26(4):221-31.
Transport of mRNA from nucleus to cytoplasm is an ATP-dependent process which occurs strictly vectorially. Because the mRNA is structurally bound during transport, mRNA transport is a "solid-state" process consisting of i) mRNA release from the nuclear matrix, ii) mRNA translocation through the nuclear pore, and iii) cytoskeletal binding. We identified and purified the following components involved in the translocation step: i) the nuclear envelope (NE) nucleoside triphosphatase (NTPase) which is stimulated by the 3'poly(A) tail of mRNA, ii) the poly(A)-recognizing mRNA carrier, iii) the NE protein kinase, and iv) the NE phosphatase. In addition, we found that an RNA helicase activity is present in NE, which also may be involved in RNA transport. Our results show that, besides poly(A), also double-stranded RNA structures may modulate RNA export. The amount of mRNA released from nuclei markedly decreases with age. Evidence is presented that this age-dependent change is caused by an impairment of polyadenylation of mRNA, hnRNA processing, release of mRNA from nuclear matrix, and translocations of mRNA from nuclear to cytoplasmic compartment (decrease in activities of NE NTPase, protein kinase, and phosphatase; decrease in poly(A)-binding affinity of mRNA carrier).
mRNA从细胞核向细胞质的转运是一个依赖ATP的过程,该过程严格按向量发生。由于mRNA在转运过程中在结构上受到束缚,mRNA转运是一个“固态”过程,包括:i)mRNA从核基质释放;ii)mRNA通过核孔易位;iii)与细胞骨架结合。我们鉴定并纯化了易位步骤中涉及的以下组分:i)受mRNA的3'聚(A)尾刺激的核被膜(NE)核苷三磷酸酶(NTPase);ii)识别聚(A)的mRNA载体;iii)NE蛋白激酶;iv)NE磷酸酶。此外,我们发现NE中存在一种RNA解旋酶活性,其也可能参与RNA转运。我们的结果表明,除了聚(A)之外,双链RNA结构也可能调节RNA输出。从细胞核释放的mRNA量随年龄显著减少。有证据表明,这种年龄依赖性变化是由mRNA的多聚腺苷酸化受损、核不均一RNA(hnRNA)加工、mRNA从核基质释放以及mRNA从细胞核向细胞质区室的易位(NE NTPase、蛋白激酶和磷酸酶活性降低;mRNA载体的聚(A)结合亲和力降低)引起的。