Hirschhorn R R, Marashi F, Baserga R, Stein J, Stein G
Biochemistry. 1984 Jul 31;23(16):3731-5. doi: 10.1021/bi00311a025.
The expression of genes coding for the four core histones (H2A, H2B, H3, and H4) was studied in tsAF8 cells. These baby hamster kidney-derived cells are a temperature-sensitive (ts) mutant of the cell cycle that arrest in G1 at the restrictive temperature. When serum-deprived tsAF8 cells are stimulated with serum, they enter the S phase at the permissive temperature of 34 degrees C, but are blocked in G1 at the nonpermissive temperature of 39.6 degrees C. Northern blot analysis using cloned human histone DNA probes detected only very low levels of histone RNA either in quiescent tsAF8 cells or in cells serum stimulated at the nonpermissive temperature for 24 h. Cellular levels of histone RNA were markedly increased in cells serum stimulated at 34 degrees C for 24 h. Temperature shift-up experiments after serum stimulation of quiescent populations showed that the amount of histone RNA was related to the number of cells that entered the S phase. Those cells that synthesized histone RNA and entered the S phase were capable of dividing. This is the first demonstration in a mammalian G1-specific ts mutant that the expression of H2A, H2B, H3, and H4 histone genes depends on the entry of cells into the S phase of the cell cycle.
在tsAF8细胞中研究了编码四种核心组蛋白(H2A、H2B、H3和H4)的基因表达。这些源自幼仓鼠肾的细胞是细胞周期的温度敏感(ts)突变体,在限制温度下停滞于G1期。当血清剥夺的tsAF8细胞受到血清刺激时,它们在34℃的允许温度下进入S期,但在39.6℃的非允许温度下被阻滞在G1期。使用克隆的人组蛋白DNA探针进行的Northern印迹分析显示,在静止的tsAF8细胞或在非允许温度下血清刺激24小时的细胞中,仅检测到非常低水平的组蛋白RNA。在34℃血清刺激24小时的细胞中,组蛋白RNA的细胞水平显著增加。对静止群体进行血清刺激后的温度上调实验表明,组蛋白RNA的量与进入S期的细胞数量有关。那些合成组蛋白RNA并进入S期的细胞能够分裂。这是在哺乳动物G1特异性ts突变体中的首次证明,即H2A、H2B、H3和H4组蛋白基因的表达取决于细胞进入细胞周期的S期。