Istituto di Chimica del Riconoscimento Molecolare, CNR, Milan, Italy.
San Raffaele Rett Research Unit, San Raffaele Scientific Institute, Milan, Italy.
Biochim Biophys Acta Gen Subj. 2018 May;1862(5):1180-1189. doi: 10.1016/j.bbagen.2018.02.005. Epub 2018 Feb 8.
Mutations in the X-linked MECP2 gene represent the main origin of Rett syndrome, causing a profound intellectual disability in females. MeCP2 is an epigenetic transcriptional regulator containing two main functional domains: a methyl-CpG binding domain (MBD) and a transcription repression domain (TRD). Over 600 pathogenic mutations were reported to affect the whole protein; almost half of missense mutations affect the MBD. Understanding the impact of these mutations on the MBD structure and interaction with DNA will foster the comprehension of their pathogenicity and possibly genotype/phenotype correlation studies. Herein, we use molecular dynamics simulations to obtain a detailed view of the dynamics of WT and mutated MBD in the presence and absence of DNA. The pathogenic mutation Y120D is used as paradigm for our studies. Further, since the Y120 residue was previously found to be a phosphorylation site, we characterize the dynamic profile of the MBD also in the presence of Y120 phosphorylation (pY120). We found that addition of a phosphate group to Y120 or mutation in aspartic acid affect domain mobility that samples an alternative conformational space with respect to the WT, leading to impaired ability to interact with DNA. Experimental assays showing a significant reduction in the binding affinity between the mutated MBD and the DNA confirmed our predictions.
X 连锁的 MECP2 基因突变是瑞特综合征的主要病因,导致女性出现严重的智力障碍。MeCP2 是一种表观遗传转录调节剂,包含两个主要的功能域:甲基化 CpG 结合域(MBD)和转录抑制域(TRD)。已报道超过 600 种致病性突变会影响整个蛋白;几乎一半的错义突变影响 MBD。了解这些突变对 MBD 结构和与 DNA 相互作用的影响,将有助于理解它们的致病性,并可能进行基因型/表型相关性研究。在此,我们使用分子动力学模拟来详细观察 WT 和突变 MBD 在存在和不存在 DNA 时的动力学。致病性突变 Y120D 被用作我们研究的范例。此外,由于 Y120 残基先前被发现是一个磷酸化位点,我们还对 MBD 存在 Y120 磷酸化(pY120)的情况下的动态特性进行了表征。我们发现,向 Y120 添加一个磷酸基团或突变天冬氨酸会影响到 MBD 的结构域的迁移率,使其能够在相对于 WT 的另一种构象空间中进行采样,从而导致与 DNA 相互作用的能力受损。实验测定表明,突变 MBD 与 DNA 之间的结合亲和力显著降低,这证实了我们的预测。