Hahm K, Cobb B S, McCarty A S, Brown K E, Klug C A, Lee R, Akashi K, Weissman I L, Fisher A G, Smale S T
Howard Hughes Medical Institute, Molecular Biology Institute, and Department of Microbiology and Immunology, UCLA School of Medicine, Los Angeles, California 90095-1662, USA.
Genes Dev. 1998 Mar 15;12(6):782-96. doi: 10.1101/gad.12.6.782.
The Ikaros gene encodes multiple protein isoforms that contribute critical functions during the development of lymphocytes and other hematopoietic cell types. The intracellular functions of Ikaros are not known, although recent studies have shown that Ikaros proteins colocalize with inactive genes and centromeric heterochromatin. In this study, Ikaros proteins were found to be components of highly stable complexes. The complexes from an immature T cell line were purified, revealing associated proteins of 70 and 30 kD. The p70 gene, named Helios, encodes two protein isoforms with zinc finger domains exhibiting considerable homology to those within Ikaros proteins. Helios and Ikaros recognize similar DNA sequences and, when overexpressed, Helios associates indiscriminately with the various Ikaros isoforms. Although Ikaros is present in most hematopoietic cells, Helios was found primarily in T cells. The relevance of the Ikaros-Helios interaction in T cells is supported by the quantitative association of Helios with a fraction of the Ikaros. Interestingly, the Ikaros-Helios complexes localize to the centromeric regions of T cell nuclei, similar to the Ikaros localization previously observed in B cells. Unlike the B cell results, however, only a fraction of the Ikaros, presumably the fraction associated with Helios, exhibited centromeric localization in T cells. These results establish immunoaffinity chromatography as a useful method for identifying Ikaros partners and suggest that Helios is a limiting regulatory subunit for Ikaros within centromeric heterochromatin.
Ikaro基因编码多种蛋白质异构体,这些异构体在淋巴细胞和其他造血细胞类型的发育过程中发挥关键作用。尽管最近的研究表明Ikaro蛋白与无活性基因和着丝粒异染色质共定位,但Ikaro的细胞内功能尚不清楚。在本研究中,发现Ikaro蛋白是高度稳定复合物的组成部分。从未成熟T细胞系中纯化出复合物,发现了70kD和30kD的相关蛋白。名为Helios的p70基因编码两种蛋白质异构体,其锌指结构域与Ikaro蛋白中的锌指结构域具有相当高的同源性。Helios和Ikaro识别相似的DNA序列,并且当过度表达时,Helios会不加区分地与各种Ikaro异构体结合。尽管Ikaro存在于大多数造血细胞中,但Helios主要存在于T细胞中。Helios与一部分Ikaro的定量结合支持了Ikaro-Helios相互作用在T细胞中的相关性。有趣的是,Ikaro-Helios复合物定位于T细胞核的着丝粒区域,类似于先前在B细胞中观察到的Ikaro定位。然而,与B细胞的结果不同,在T细胞中只有一部分Ikaro(可能是与Helios相关的部分)表现出着丝粒定位。这些结果确立了免疫亲和色谱法作为鉴定Ikaro伙伴的有用方法,并表明Helios是着丝粒异染色质中Ikaro的一种限制性调节亚基。