Mapping the micro-proteome of the nuclear lamina and lamina-associated domains.
Mapping the micro-proteome of the nuclear lamina and lamina-associated domains.
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DOI：
10.26508/lsa.202000774
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发表时间：
2021-05
影响因子：
4.4
通讯作者：
Reddy KL
中科院分区：
文献类型：
--
作者：
Wong X;Cutler JA;Hoskins VE;Gordon M;Madugundu AK;Pandey A;Reddy KL
核纤层为细胞核提供结构，并作为细胞骨架和称为LAD的大异染色质结构域之间的界面。本研究描述了这种LAD/椎板界面的微蛋白质组。核纤层是一种蛋白质纤维网络，通过将蛋白质和DNA的大结构域（所谓的纤层相关结构域（LAD））束缚到核的外周来提供结构和基因调控功能。LAD是哺乳动物基因组的很大一部分，部分是由于它们与核外围的结合而受到抑制。对LAD的发生和维持以及参与这些功能的蛋白质知之甚少。在努力确定蛋白质，居住在核周边和潜在的相互作用与LAD，我们已经采取了双管齐下的方法。首先，我们对内核膜结合的LAP 2 β进行了相互作用组分析，以进一步表征核纤层蛋白质组。为了实现这一目标，我们利用了BioID系统，该系统以前已成功地用于表征核纤层蛋白质组。第二，我们已经建立了一个系统，以确定蛋白质结合的LAD通过开发一个染色质导向的BioID系统。我们将BioID系统与结合活细胞中的LAD的m6 A-示踪剂系统相结合，以鉴定LAD近端和核纤层蛋白。在结合这些数据集，我们已经进一步的特点，在核纤层的蛋白质网络，确定了推定的LAD近端蛋白质，并发现了几个蛋白质，似乎与两个微蛋白质组接口。重要的是，在这项研究中鉴定了几种对LAD功能至关重要的蛋白，包括与H3 K9甲基化相关的异染色质调节蛋白。
The nuclear lamina provides structure to the nucleus and serves as an interface between the cytoskeleton and large heterochromatin domains called LADs. This study describes the microproteome of this LAD/lamina interface. The nuclear lamina is a proteinaceous network of filaments that provide both structural and gene regulatory functions by tethering proteins and large domains of DNA, the so-called lamina-associated domains (LADs), to the periphery of the nucleus. LADs are a large fraction of the mammalian genome that are repressed, in part, by their association to the nuclear periphery. The genesis and maintenance of LADs is poorly understood as are the proteins that participate in these functions. In an effort to identify proteins that reside at the nuclear periphery and potentially interact with LADs, we have taken a two-pronged approach. First, we have undertaken an interactome analysis of the inner nuclear membrane bound LAP2β to further characterize the nuclear lamina proteome. To accomplish this, we have leveraged the BioID system, which previously has been successfully used to characterize the nuclear lamina proteome. Second, we have established a system to identify proteins that bind to LADs by developing a chromatin-directed BioID system. We combined the BioID system with the m6A-tracer system which binds to LADs in live cells to identify both LAD proximal and nuclear lamina proteins. In combining these datasets, we have further characterized the protein network at the nuclear lamina, identified putative LAD proximal proteins and found several proteins that appear to interface with both micro-proteomes. Importantly, several proteins essential for LAD function, including heterochromatin regulating proteins related to H3K9 methylation, were identified in this study.
影响因子：
1.8
作者：
Hsu FH;Chen HI;Tsai MH;Lai LC;Huang CC;Tu SH;Chuang EY;Chen Y
通讯作者：
Chen Y