Stoldt Stefan, Wenzel Dirk, Schulze Ekkehard, Doenecke Detlef, Happel Nicole
Institute for Biochemistry and Molecular Cell Biology, University of Göttingen, Humboldtallee 23, 37073 Göttingen, Germany.
Biol Cell. 2007 Oct;99(10):541-52. doi: 10.1042/bc20060117.
H1 histones are a protein family comprising several subtypes. Although specific functions of the individual subtypes could not be determined so far, differential roles are indicated by varied nuclear distributions as well as differential expression patterns of the H1 subtypes. Although the group of replication-dependent H1 subtypes is synthesized during S phase, the replacement H1 subtype, H1 degrees , is also expressed in a replication-independent manner in non-proliferating cells. Recently we showed, by protein biochemical analysis, that the ubiquitously expressed subtype H1x is enriched in the micrococcal nuclease-resistant part of chromatin and that, although it shares common features with H1 degrees , its expression is differentially regulated, since, in contrast to H1 degrees , growth arrest or induction of differentiation did not induce an accumulation of H1x.
In the present study, we show that H1x exhibits a cell-cycle-dependent change of its nuclear distribution. This H1 subtype showed a nucleolar accumulation during the G(1) phase, and it was evenly distributed in the nucleus during S phase and G(2). Immunocytochemical analysis of the intranucleolar distribution of H1x indicated that it is located mainly in the condensed nucleolar chromatin. In addition, we demonstrate that the amount of H1x protein remained nearly unchanged during S phase progression, which is in contrast to the replication-dependent subtypes.
These results suggest that the differential localization of H1x provides a mechanism for a control of H1x activity by means of shuttling between nuclear subcompartments instead of a controlled turnover of the protein.
H1组蛋白是一个包含多种亚型的蛋白质家族。尽管目前尚未确定各个亚型的具体功能,但不同的核分布以及H1亚型的差异表达模式表明了它们具有不同的作用。虽然依赖复制的H1亚型在S期合成,但替代型H1亚型H1°在非增殖细胞中也以不依赖复制的方式表达。最近,我们通过蛋白质生化分析表明,普遍表达的亚型H1x在染色质的微球菌核酸酶抗性部分中富集,并且尽管它与H1°具有共同特征,但其表达受到差异调节,因为与H1°不同,生长停滞或诱导分化不会导致H1x积累。
在本研究中,我们表明H1x的核分布呈现细胞周期依赖性变化。这种H1亚型在G1期表现为核仁积累,在S期和G2期均匀分布于细胞核中。对H1x在核仁内分布的免疫细胞化学分析表明,它主要位于浓缩的核仁染色质中。此外,我们证明在S期进程中H1x蛋白的量几乎保持不变,这与依赖复制的亚型不同。
这些结果表明,H1x的差异定位提供了一种机制,通过在核亚区室之间穿梭来控制H1x的活性,而不是通过控制蛋白质的周转。