Sekiguchi T, Nakashima T, Hayashida T, Kuraoka A, Hashimoto S, Tsuchida N, Shibata Y, Hunter T, Nishimoto T
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka, Japan.
Exp Cell Res. 1995 Jun;218(2):490-8. doi: 10.1006/excr.1995.1183.
A temperature-sensitive (ts) mutant of the BHK21 cell line derived from golden hamsters, tsBN462 has a mutation in the gene encoding the largest subunit of the TFIID complex, TAFII250/p230/CCG1, and arrests in the G1 phase at the nonpermissive temperature, 39.5 degrees C. We found that tsBN462 cells underwent apoptosis following growth arrest at 39.5 degrees C, suggesting a role for CCG1 as a repressor of apoptosis. By electron microscopic observation, tsBN462 cells at 39.5 degrees C showed characteristic features of apoptosis. Apoptosis was not suppressed by expression of Bc1-2 or the adenovirus E1B 19 kDa protein. Cell death was suppressed completely by expression of wild-type CCG1 and partially by wild-type p53, a growth suppressor protein. Cell cycle arrest induced by p53 may help survival of tsBN462 cells at 39.5 degrees C. Apoptosis was accelerated in SV40 large T antigen-transformed tsBN462 cells at 39.5 degrees C where SV40 large T antigen formed a complex with p53, implying that the apoptosis of tsBN462 cells at 39.5 degrees C occurred in a p53-independent manner. Our results suggest that CCG1/TAFII250 is required for the expression of factors regulating apoptosis.
tsBN462是一种源自金黄地鼠的BHK21细胞系的温度敏感(ts)突变体,其编码TFIID复合物最大亚基TAFII250/p230/CCG1的基因发生了突变,并在非允许温度39.5℃时停滞于G1期。我们发现,tsBN462细胞在39.5℃生长停滞之后会发生凋亡,这表明CCG1作为凋亡抑制因子发挥作用。通过电子显微镜观察,处于39.5℃的tsBN462细胞呈现出凋亡的特征性表现。凋亡并未被Bc1-2或腺病毒E1B 19 kDa蛋白的表达所抑制。细胞死亡被野生型CCG1的表达完全抑制,并被生长抑制蛋白野生型p53部分抑制。p53诱导的细胞周期停滞可能有助于tsBN462细胞在39.5℃存活。在39.5℃时,SV40大T抗原转化的tsBN462细胞中的凋亡加速,此时SV40大T抗原与p53形成复合物,这意味着tsBN462细胞在39.5℃时的凋亡以p53非依赖的方式发生。我们的结果表明,CCG1/TAFII250是调节凋亡的因子表达所必需的。