Lee Y M, Lee S, Lee E, Shin H, Hahn H, Choi W, Kim W
Institute for Medical Sciences, School of Medicine, Ajou University, Suwon 442-749, South Korea.
Biochem J. 2001 Dec 15;360(Pt 3):549-56. doi: 10.1042/0264-6021:3600549.
In a previous study, we identified the human counterpart of murine kinesin superfamily member 4 (KIF4), a microtubule-based motor protein [Oh, Hahn, Torrey, Shin, Choi, Lee, Morse and Kim (2000) Biochim. Biophys. Acta 1493, 219-224]. As an initial step to understand the function(s) of human KIF4, its subcellular localization in HeLa cells was examined by using immunocytochemical and subcellular fractionation methods, and it was found that most KIF4 is localized in the nucleus. Since murine KIF4 is known to transport cytoplasmic vesicles, dominant nuclear localization of the human counterpart was somewhat surprising. Subsequent subnuclear fractionation revealed predominant association of KIF4 with the nuclear matrix. These results clearly indicate that human KIF4 is, at least, a nuclear protein. In further confirmation of this conclusion, the hexapeptide PKLRRR (amino acids 773-778) in the molecule was found to function as a nuclear localization signal. During the mitotic phase of the cell cycle, human KIF4 was associated with the chromosomes, suggesting that human KIF4 might be a microtubule-based mitotic motor, with DNA as its cargo.
在先前的一项研究中,我们鉴定出了小鼠驱动蛋白超家族成员4(KIF4)的人类对应物,它是一种基于微管的驱动蛋白[Oh, Hahn, Torrey, Shin, Choi, Lee, Morse和Kim(2000年),《生物化学与生物物理学报》1493, 219 - 224]。作为了解人类KIF4功能的第一步,我们通过免疫细胞化学和亚细胞分级分离方法检测了它在HeLa细胞中的亚细胞定位,发现大多数KIF4定位于细胞核。由于已知小鼠KIF4可运输细胞质囊泡,人类对应物的主要核定位有些令人惊讶。随后的细胞核亚分级分离显示KIF4主要与核基质相关联。这些结果清楚地表明人类KIF4至少是一种核蛋白。为进一步证实这一结论,发现该分子中的六肽PKLRRR(氨基酸773 - 778)起到核定位信号的作用。在细胞周期的有丝分裂期,人类KIF4与染色体相关联,这表明人类KIF4可能是一种基于微管的有丝分裂驱动蛋白,以DNA为其运载物。