Balasundaram D, Benedik M J, Morphew M, Dang V D, Levin H L
Laboratory of Eukaryotic Gene Regulation, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA.
Mol Cell Biol. 1999 Aug;19(8):5768-84. doi: 10.1128/MCB.19.8.5768.
The long terminal repeat (LTR)-containing retrotransposon Tf1 propagates within the fission yeast Schizosaccharomyces pombe as the result of several mechanisms that are typical of both retrotransposons and retroviruses. To identify host factors that contribute to the transposition process, we mutagenized cultures of S. pombe and screened them for strains that were unable to support Tf1 transposition. One such strain contained a mutation in a gene we named nup124. The product of this gene contains 11 FXFG repeats and is a component of the nuclear pore complex. In addition to the reduced levels of Tf1 transposition, the nup124-1 allele caused a significant reduction in the nuclear localization of Tf1 Gag. Surprisingly, the mutation in nup124-1 did not cause any reduction in the growth rate, the nuclear localization of specific nuclear localization signal-containing proteins, or the cytoplasmic localization of poly(A) mRNA. A two-hybrid analysis and an in vitro precipitation assay both identified an interaction between Tf1 Gag and the N terminus of Nup124p. These results provide evidence for an unusual mechanism of nuclear import that relies on a direct interaction between a nuclear pore factor and Tf1 Gag.
含有长末端重复序列(LTR)的逆转座子Tf1在裂殖酵母粟酒裂殖酵母中传播,这是逆转座子和逆转录病毒共有的几种典型机制作用的结果。为了鉴定有助于转座过程的宿主因子,我们对粟酒裂殖酵母培养物进行诱变,并筛选出不能支持Tf1转座的菌株。其中一个这样的菌株在我们命名为nup124的基因中发生了突变。该基因的产物含有11个FXFG重复序列,是核孔复合体的一个组成部分。除了Tf1转座水平降低外,nup124-1等位基因还导致Tf1 Gag的核定位显著降低。令人惊讶的是,nup124-1中的突变并未导致生长速率、含特定核定位信号蛋白的核定位或聚腺苷酸mRNA的细胞质定位有任何降低。双杂交分析和体外沉淀试验均鉴定出Tf1 Gag与Nup124p的N末端之间存在相互作用。这些结果为一种不寻常的核输入机制提供了证据,该机制依赖于核孔因子与Tf1 Gag之间的直接相互作用。