Bennett C F, Yeoman L C
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030.
Biochem J. 1987 Oct 1;247(1):109-12. doi: 10.1042/bj2470109.
We have previously shown that a 30 kDa DNA-binding protein isolated from rat cell nuclei exhibits the chemical and immunological properties of glutathione S-transferase Yb subunits [Bennett, Spector & Yeoman (1986) J. Cell Biol. 102, 600-609]. It was of interest, therefore, to determine whether Yb subunits isolated from rat liver nuclei would return to nuclear fractions upon reintroduction to cell cytoplasms via red-blood-cell-mediated fusion. Labelled Yb subunits were associated with nuclear fractions 60 min after cell fusion. The microinjected protein remained associated with the nuclei for 18 h and was not extractable with low-salt washes. In addition, injected Yb subunits were found to equally distribute between extractable (56%) and residual (44%) nuclear fractions. These experiments demonstrate that glutathione S-transferase Yb subunits isolated from nuclei rapidly translocate to nuclei upon reintroduction into cell cytoplasms.
我们之前已经表明,从大鼠细胞核中分离出的一种30 kDa的DNA结合蛋白具有谷胱甘肽S-转移酶Yb亚基的化学和免疫学特性[贝内特、斯佩克特和约曼(1986年)《细胞生物学杂志》102卷,600 - 609页]。因此,确定从大鼠肝细胞核中分离出的Yb亚基通过红细胞介导的融合重新引入细胞质后是否会返回核组分是很有意义的。细胞融合60分钟后,标记的Yb亚基与核组分相关联。显微注射的蛋白与细胞核结合长达18小时,并且不能用低盐洗涤液提取。此外,发现注射的Yb亚基在可提取的(56%)和残留的(44%)核组分之间均匀分布。这些实验表明,从细胞核中分离出的谷胱甘肽S-转移酶Yb亚基在重新引入细胞质后会迅速转运到细胞核中。