Abstract
Sec13p has been known as an endoplasmic reticulum-Golgi transport protein. Recently, it has also been shown to be required for the formation of septation in the fission yeast Schizosaccharomyces pombe. In the present study, we focused on the role of a human homolog of Saccharomyces cerevisiae SEC13, Sec13 protein during mitosis in U2OS cells. We found that the expression of Sec13 was constant throughout the cell cycle, and localized to the kinetochores at metaphase during mitosis. By using green fluorescent protein technology, we observed that Sec13 is required for evasion of mitotic arrest in response to spindle damage, leading to G1-like phase and apoptotic cell death. In addition, cells expressing exogenous Sec13 showed giant nuclei compared to endogenous ones in the absence of nocodazole. These results demonstrate that Sec13 is involved in the regulation of the metaphase/anaphase transition and may be functionally associated with mitotic machinery to maintain genomic stability during mitosis.
Highlights
Yeast Sec13p is a component of the yeast coat protein II (COP II) complex mediating the process of vesicle budding from endoplasmic reticulum (ER) to Golgi apparatus (Pryer et al, 1993; Lederkremer et al, 2001)
In the present study, we focused on whether Sec[13] is involved in regulating mitotic progression in mammalian cells
Human osteosarcoma U2OS cells were cultured in Dubelcco's modified essential medium (DMEM) supplemented with 10% fetal bovine serum, antibiotics and incubated in humidified incubator at 37oC with 5% CO2
Summary
Yeast Sec13p is a component of the yeast coat protein II (COP II) complex mediating the process of vesicle budding from endoplasmic reticulum (ER) to Golgi apparatus (Pryer et al, 1993; Lederkremer et al, 2001). Like yeast Sec13p, human Sec[13] protein is essential for the transport of ER to Golgi and is a component of mammalian COP II complex as a coat and adapter protein (Tang et al, 1997; Lederkremer et al, 2001). The subcellular localization of Sec[13] has been observed on the membranes of the nuclear pore in yeast and mammalian cells (Siniossoglou et al, 1996). Nup[107] localizes at kinetochores during mitosis (Belgarch et al, 2001). In yeast, Mad[1] and Mad[2], components of the spindle checkpoint, localize on the nuclear membrane during interphase and bind to the NPC complex (Campbell et al, 2001; Iouk et al, 2002). In the present study, we focused on whether Sec[13] is involved in regulating mitotic progression in mammalian cells
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