Nielsen A L, Ortiz J A, You J, Oulad-Abdelghani M, Khechumian R, Gansmuller A, Chambon P, Losson R
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP/Collège de France, BP 163, 67404 Illkirch-Cedex, France.
EMBO J. 1999 Nov 15;18(22):6385-95. doi: 10.1093/emboj/18.22.6385.
Mammalian TIF1alpha and TIF1beta (KAP-1/KRIP-1) are related transcriptional intermediary factors that possess intrinsic silencing activity. TIF1alpha is believed to be a euchromatic target for liganded nuclear receptors, while TIF1beta may serve as a co-repressor for the large family of KRAB domain-containing zinc finger proteins. Here, we report an association of TIF1beta with both heterochromatin and euchromatin in interphase nuclei. Co-immunoprecipitation of nuclear extracts shows that endogenous TIF1beta, but not TIF1alpha, is associated with members of the heterochromatin protein 1 (HP1) family. However, in vitro, both TIF1alpha and TIF1beta interact with and phosphorylate the HP1 proteins. This interaction involves a conserved amino acid motif, which is critical for the silencing activity of TIF1beta but not TIF1alpha. We further show that trichostatin A, an inhibitor of histone deacetylases, can interfere with both TIF1 and HP1 silencing. The silencing activity of TIF1alpha appears to result chiefly from histone deacetylation, whereas that of TIF1beta may be mediated via both HP1 binding and histone deacetylation.
哺乳动物的TIF1α和TIF1β(KAP-1/KRIP-1)是相关的转录中介因子,具有内在的沉默活性。TIF1α被认为是配体核受体的常染色质靶点,而TIF1β可能作为包含KRAB结构域的锌指蛋白大家族的共抑制因子。在此,我们报道了TIF1β在间期细胞核中与异染色质和常染色质均有关联。对核提取物进行的共免疫沉淀显示,内源性TIF1β而非TIF1α与异染色质蛋白1(HP1)家族成员相关联。然而,在体外,TIF1α和TIF1β均与HP1蛋白相互作用并使其磷酸化。这种相互作用涉及一个保守的氨基酸基序,这对TIF1β的沉默活性至关重要,但对TIF1α并非如此。我们进一步表明,组蛋白脱乙酰酶抑制剂曲古抑菌素A可干扰TIF1和HP1的沉默作用。TIF1α的沉默活性似乎主要源于组蛋白去乙酰化,而TIF1β的沉默活性可能通过HP1结合和组蛋白去乙酰化两者介导。