Barrier-to-autointegration factor proteome reveals chromatin-regulatory partners.
Barrier-to-autointegration factor proteome reveals chromatin-regulatory partners.
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DOI：
10.1371/journal.pone.0007050
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发表时间：
2009-09-16
期刊：
影响因子：
3.7
通讯作者：
Wilson KL
中科院分区：
文献类型：
--
作者：
Montes de Oca R;Shoemaker CJ;Gucek M;Cole RN;Wilson KL
核核纤层蛋白丝和相关蛋白质形成多细胞动物中转录、复制、染色质组织和表观遗传调控所需的核骨架（“纤层”）网络。核纤层缺陷导致人类疾病（“核纤层病”），并与衰老有关。自整合屏障因子（BAF）是核纤层的一种移动的必需组分，其直接与组蛋白、核纤层蛋白和LEM结构域蛋白（包括内核膜蛋白emerin）结合，并且在染色质结构、有丝分裂和基因调控中起作用。为了理解BAF的作用机制，使用BAF缀合的珠从HeLa细胞核裂解物中亲和纯化BAF相关蛋白，并通过串联质谱法鉴定或使用iTRAQ方法独立鉴定和定量。我们回收了A-和B-型核纤层蛋白和核心组蛋白，所有已知的都直接结合BAF，加上四种人转录因子（p15，NonO，LEDGF），疾病相关蛋白（例如，Huntingtin，Treacle）和几种调节染色质的蛋白质和酶。与内源性BAF的关联通过来自HeLa细胞的七种候选物的共免疫沉淀独立地验证，所述候选物包括DIFFEEM、聚（ADP-核糖）聚合酶1（PARP 1）、视网膜母细胞瘤结合蛋白4（RBBP 4）、损伤特异性DNA结合蛋白1（DDB 1）和DDB 2。有趣的是，内源性BAF和emerin每个与DDB 2和CUL 4A在紫外线和时间依赖性的方式，表明BAF和emerin基因组的完整性有动态的作用，并可能有助于耦合DNA损伤反应的核纤层网络。我们得出结论，这种蛋白质组是一个丰富的来源的候选合作伙伴BAF和潜在的A型和B型核纤层蛋白，这可能会揭示如何染色质调控和基因组的完整性与核结构。
Nuclear lamin filaments and associated proteins form a nucleoskeletal (“lamina”) network required for transcription, replication, chromatin organization and epigenetic regulation in metazoans. Lamina defects cause human disease (“laminopathies”) and are linked to aging. Barrier-to-autointegration factor (BAF) is a mobile and essential component of the nuclear lamina that binds directly to histones, lamins and LEM-domain proteins, including the inner nuclear membrane protein emerin, and has roles in chromatin structure, mitosis and gene regulation. To understand BAF's mechanisms of action, BAF associated proteins were affinity-purified from HeLa cell nuclear lysates using BAF-conjugated beads, and identified by tandem mass spectrometry or independently identified and quantified using the iTRAQ method. We recovered A- and B-type lamins and core histones, all known to bind BAF directly, plus four human transcription factors (Requiem, NonO, p15, LEDGF), disease-linked proteins (e.g., Huntingtin, Treacle) and several proteins and enzymes that regulate chromatin. Association with endogenous BAF was independently validated by co-immunoprecipitation from HeLa cells for seven candidates including Requiem, poly(ADP-ribose) polymerase 1 (PARP1), retinoblastoma binding protein 4 (RBBP4), damage-specific DNA binding protein 1 (DDB1) and DDB2. Interestingly, endogenous BAF and emerin each associated with DDB2 and CUL4A in a UV- and time-dependent manner, suggesting BAF and emerin have dynamic roles in genome integrity and might help couple DNA damage responses to the nuclear lamina network. We conclude this proteome is a rich source of candidate partners for BAF and potentially also A- and B-type lamins, which may reveal how chromatin regulation and genome integrity are linked to nuclear structure.
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DOI：
10.1038/2345
发表时间：
1998-10-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
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通讯作者：
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影响因子：
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