Meergans T, Albig W, Doenecke D
Institut für Biochemie und Molekulare Zellbiologie, Abteilung Molekularbiologie, Universität Göttingen, Germany.
Eur J Biochem. 1998 Sep 1;256(2):436-46. doi: 10.1046/j.1432-1327.1998.2560436.x.
In man, the H1 class of histones consists of seven different isoforms, termed H1.1-H1.5, H1t and H1o. Analysis of the promoters of the respective genes reveals that all seven H1 gene promoters share conserved sequence elements: a TATA box at around position -25 (relative to the transcription start site), a CCAAT motif at about position -50 (except in the H1 promoter), an H1-box (AAACACA) around position -110 (except in the H1.1 promoter), and the highly conserved motif TGTGT/CTA (TG-box or CH1UE) at around nucleotide position -450 (except in the H1.1 promoter). Analysis of the H1.3 gene promoter was carried out with reporter gene assays (using the luciferase gene as a reporter gene) including stepwise deletion and site-directed mutagenesis. In addition, electrophoretic mobility-shift assays were carried out for the analysis of protein/DNA interactions at conserved promoter motifs. Mutation analysis indicates that the CH1UE motif is involved in mediating the S-phase-dependent expression of the H1.3 gene. Comparison of H1 promoters shows that the CCAAT-box is extended in each case by CA. Mutational analysis indicates that only the CCAATCA heptanucleotide, but not just the CCAAT sequence mediates the effect of this element in H1 gene promoters.
在人类中,组蛋白H1家族由七种不同的亚型组成,分别称为H1.1 - H1.5、H1t和H1o。对各个基因启动子的分析表明,所有七个H1基因启动子都共享保守的序列元件:在大约-25位置(相对于转录起始位点)有一个TATA盒,在大约-50位置有一个CCAAT基序(H1启动子除外),在大约-110位置有一个H1盒(AAACACA)(H1.1启动子除外),以及在核苷酸位置大约-450处有高度保守的基序TGTGT/CTA(TG盒或CH1UE)(H1.1启动子除外)。使用报告基因测定法(以荧光素酶基因为报告基因),包括逐步缺失和定点诱变,对H1.3基因启动子进行了分析。此外,还进行了电泳迁移率变动分析,以分析保守启动子基序处的蛋白质/DNA相互作用。突变分析表明,CH1UE基序参与介导H1.3基因的S期依赖性表达。H1启动子的比较表明,每种情况下CCAAT盒都由CA延伸。突变分析表明,只有CCAATCA七核苷酸,而不仅仅是CCAAT序列介导了该元件在H1基因启动子中的作用。