Davey J, Dimmock N J, Colman A
Cell. 1985 Mar;40(3):667-75. doi: 10.1016/0092-8674(85)90215-6.
Influenza virus nucleoprotein (NP), synthesized in Xenopus oocytes after injection of cloned NP cDNA, enters and accumulates in the nucleus. We have used in vitro mutagenesis of this cDNA to study the cellular distribution of mutated NP polypeptides. Mutants lacking amino acids 327-345 of wild-type NP enter the nucleus but do not accumulate there to the same extent as the wild-type protein, suggesting that this region has a role in nuclear accumulation. This possibility is further strengthened by similar studies involving the production of fusion proteins in which various amino-terminal sequences of the NP gene are fused to the complete chimpanzee alpha 1-globin sequence: when globin cDNA was injected into and expressed in oocytes the protein remains exclusively in the cytosol; however, when the globin cDNA is fused to a portion of NP cDNA that includes the region encoding amino acids 327-345, the resulting fusion protein enters and accumulates in the nucleus. Fusion proteins lacking this region of the NP enter but do not accumulate in the nucleus.
将克隆的核蛋白(NP)cDNA注射到非洲爪蟾卵母细胞中合成的流感病毒核蛋白进入细胞核并在其中积累。我们利用该cDNA的体外诱变来研究突变NP多肽的细胞分布。缺乏野生型NP氨基酸327 - 345的突变体进入细胞核,但在细胞核中的积累程度不如野生型蛋白,这表明该区域在核积累中起作用。涉及产生融合蛋白的类似研究进一步强化了这种可能性,其中NP基因的各种氨基末端序列与完整的黑猩猩α1 - 珠蛋白序列融合:当将珠蛋白cDNA注射到卵母细胞中并在其中表达时,该蛋白仅保留在细胞质中;然而,当珠蛋白cDNA与NP cDNA的一部分融合,该部分包括编码氨基酸327 - 345的区域时,产生的融合蛋白进入细胞核并在其中积累。缺乏NP该区域的融合蛋白进入细胞核但不在其中积累。