Zasloff M
Proc Natl Acad Sci U S A. 1983 Nov;80(21):6436-40. doi: 10.1073/pnas.80.21.6436.
The mechanism by which a tRNA molecule is delivered from the nucleus of a cell to the cytoplasm has been studied in the Xenopus laevis oocyte utilizing nuclear microinjection and manual microdissection techniques. tRNA nuclear transport in this cell resembles a carrier-mediated translocation process rather than diffusion through a simple pore or channel. tRNA transport is saturable by tRNA, with a maximal rate measured to be about 190 X 10(7) molecules per min per nucleus (21 degrees C) in the mature oocyte. Competitive inhibition between two different tRNA species can be demonstrated, suggesting that many tRNA species share a common carrier system. tRNA nuclear transport is sharply dependent on temperature, with an optimal rate observed at 31 degrees C. A single G-to-U substitution at position 57 in the vertebrate tRNAMeti molecule reduces the transport rate of this tRNA by a factor of about 20, implicating this highly conserved region of the tRNA molecule (loop IV) as critical for recognition by the transport mechanism. On morphologic grounds I propose that ribosome-like components surrounding the nuclear pore may function as the tRNA translocation "motor." The tRNA nuclear transport mechanism represents a distinctly eukaryotic process and a site of potential control over cell growth and proliferation.
利用核显微注射和手工显微切割技术,在非洲爪蟾卵母细胞中研究了tRNA分子从细胞核转运至细胞质的机制。该细胞中的tRNA核转运类似于载体介导的转运过程,而非通过简单的孔道或通道进行扩散。tRNA转运可被tRNA饱和,在成熟卵母细胞中,测得的最大转运速率约为每分钟每个细胞核190×10⁷个分子(21℃)。可以证明两种不同tRNA之间存在竞争性抑制,这表明许多tRNA共用一个共同的载体系统。tRNA核转运强烈依赖于温度,在31℃时观察到最佳转运速率。脊椎动物tRNAMeti分子第57位的单个G到U替换使该tRNA的转运速率降低约20倍,这表明tRNA分子的这个高度保守区域(环IV)对于转运机制的识别至关重要。基于形态学原因,我提出核孔周围类似核糖体的成分可能作为tRNA转运的“马达”。tRNA核转运机制代表了一个独特的真核过程,也是对细胞生长和增殖进行潜在调控的位点。