Haas Jana, Bloesel Daniel, Bacher Susanne, Kracht Michael, Schmitz M Lienhard
Institute of Biochemistry, Justus-Liebig-University, Giessen, Germany.
Member of the German Center for Lung Research, Giessen, Germany.
Front Cell Dev Biol. 2020 Sep 1;8:852. doi: 10.3389/fcell.2020.00852. eCollection 2020.
The protein kinase homeodomain-interacting protein kinase 2 (HIPK2) plays an important role in development and in the response to external cues. The kinase associates with an exceptionally large number of different transcription factors and chromatin regulatory proteins to direct distinct gene expression programs. In order to investigate the function of HIPK2 for chromatin compaction, HIPK2 was fused to the DNA-binding domains of Gal4 or LacI, thus allowing its specific targeting to binding sites for these transcription factors that were integrated in specific chromosome loci. Tethering of HIPK2 resulted in strong decompaction of euchromatic and heterochromatic areas. HIPK2-mediated heterochromatin decondensation started already 4 h after its chromatin association and required the functionality of its SUMO-interacting motif. This process was paralleled by disappearance of the repressive H3K27me3 chromatin mark, recruitment of the acetyltransferases CBP and p300 and increased histone acetylation at H3K18 and H4K5. HIPK2-mediated chromatin decompaction was strongly inhibited in the presence of a CBP/p300 inhibitor and completely blocked by the BET inhibitor JQ1, consistent with a causative role of acetylations for this process. Chromatin tethering of HIPK2 had only a minor effect on basal transcription, while it strongly boosted estrogen-triggered gene expression by acting as a transcriptional cofactor.
蛋白激酶同源结构域相互作用蛋白激酶2(HIPK2)在发育过程及对外界信号的应答中发挥着重要作用。该激酶与大量不同的转录因子及染色质调节蛋白相关联,以指导不同的基因表达程序。为了研究HIPK2在染色质压缩方面的功能,将HIPK2与Gal4或LacI的DNA结合结构域融合,从而使其能够特异性靶向整合于特定染色体位点的这些转录因子的结合位点。HIPK2的 tethering 导致常染色质和异染色质区域强烈解压缩。HIPK2介导的异染色质去浓缩在其与染色质结合后4小时就已开始,并且需要其SUMO相互作用基序的功能。这一过程伴随着抑制性H3K27me3染色质标记的消失、乙酰转移酶CBP和p300的募集以及H3K18和H4K5处组蛋白乙酰化增加。在存在CBP/p300抑制剂的情况下,HIPK2介导的染色质解压缩受到强烈抑制,并且被BET抑制剂JQ1完全阻断,这与乙酰化在此过程中的因果作用一致。HIPK2的染色质 tethering 对基础转录只有轻微影响,而它作为转录辅因子强烈促进雌激素触发的基因表达。