The DNA binding CXC domain of MSL2 is required for faithful targeting the Dosage Compensation Complex to the X chromosome.
The DNA binding CXC domain of MSL2 is required for faithful targeting the Dosage Compensation Complex to the X chromosome.
复制标题
DOI：
10.1093/nar/gkq026
复制
发表时间：
2010-06
影响因子：
14.9
通讯作者：
Becker PB
中科院分区：
文献类型：
--
作者：
Fauth T;Müller-Planitz F;König C;Straub T;Becker PB
果蝇的剂量补偿涉及剂量补偿复合物（DCC）对雄性 X 染色体的选择性靶向以及大多数基因的协调约 2 倍激活。 DCC 区分 X 染色体和常染色体的原理尚不清楚。靶向可能涉及 DNA 序列元件，其组合或富集标记 X 染色体。表征“染色体进入位点”或“高亲和力位点”的 DNA 序列可能具有这样的功能。然而，迄今为止，尚未在 DCC 亚基内鉴定出可以解释序列信息的 DNA 结合域。早期遗传学研究表明，MSL1 和 MSL2 可识别体内高亲和力位点 (HAS)，但尚未研究这些 DCC 亚基与 DNA 的直接相互作用。我们现在证明重组 MSL2 通过其 CXC 结构域以低纳摩尔亲和力直接结合 DNA。 MSL2 或 MSL2-MSL1 复合物的 DNA 结合在体外不会区分不同序列，但在体内报告基因测定中，表明存在未知的选择性辅因子。报告基因检测和 GFP 融合蛋白的定位证实了 CXC 结构域对体内 DCC 靶向的重要贡献。
Dosage compensation in Drosophila melanogaster involves the selective targeting of the male X chromosome by the dosage compensation complex (DCC) and the coordinate, ∼2-fold activation of most genes. The principles that allow the DCC to distinguish the X chromosome from the autosomes are not understood. Targeting presumably involves DNA sequence elements whose combination or enrichment mark the X chromosome. DNA sequences that characterize ‘chromosomal entry sites’ or ‘high-affinity sites’ may serve such a function. However, to date no DNA binding domain that could interpret sequence information has been identified within the subunits of the DCC. Early genetic studies suggested that MSL1 and MSL2 serve to recognize high-affinity sites (HAS) in vivo, but a direct interaction of these DCC subunits with DNA has not been studied. We now show that recombinant MSL2, through its CXC domain, directly binds DNA with low nanomolar affinity. The DNA binding of MSL2 or of an MSL2–MSL1 complex does not discriminate between different sequences in vitro, but in a reporter gene assay in vivo, suggesting the existence of an unknown selectivity cofactor. Reporter gene assays and localization of GFP-fusion proteins confirm the important contribution of the CXC domain for DCC targeting in vivo.
登录
查看更多内容
影响因子：
64.5
作者：
Alekseyenko AA;Peng S;Larschan E;Gorchakov AA;Lee OK;Kharchenko P;McGrath SD;Wang CI;Mardis ER;Park PJ;Kuroda MI
通讯作者：
Kuroda MI
影响因子：
10.5
作者：
Hamada, FN;Park, PJ;Kuroda, MI
通讯作者：
Kuroda, MI
影响因子：
5.3
作者：
Li, Fang;Schlemann, Anja H.;Scott, Maxwell J.
通讯作者：
Scott, Maxwell J.
影响因子：
5.3
作者：
Bell, Oliver;Conrad, Thomas;Schuebeler, Dirk
通讯作者：
Schuebeler, Dirk
影响因子：
16
作者：
Buscaino, A;Köcher, T;Akhtar, A
通讯作者：
Akhtar, A