Autophagy receptor NDP52 alters DNA conformation to modulate RNA polymerase II transcription
Autophagy receptor NDP52 alters DNA conformation to modulate RNA polymerase II transcription
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DOI：
10.1101/2022.02.01.478690
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发表时间：
2022-02
影响因子：
16.6
通讯作者：
Ália dos Santos;Daniel E. Rollins;Yukti Hari-Gupta;Hannah Reed;M. Du;Sabrina Yong Zi Ru;Kseniia Pidlisna;Ane Stranger;Faeeza Lorgat;Ian Brown;K. Howland;J. Aaron;Lin Wang;P. Ellis;T. Chew;M. Martin-Fernandez;Alice L. B. Pyne;C. Toseland
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文献类型：
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作者：
Ália dos Santos;Daniel E. Rollins;Yukti Hari-Gupta;Hannah Reed;M. Du;Sabrina Yong Zi Ru;Kseniia Pidlisna;Ane Stranger;Faeeza Lorgat;Ian Brown;K. Howland;J. Aaron;Lin Wang;P. Ellis;T. Chew;M. Martin-Fernandez;Alice L. B. Pyne;C. Toseland
NDP52 是一种自噬受体，参与入侵病原体和受损细胞器的识别和降解。尽管NDP52首先在细胞核中被鉴定并在整个细胞中表达，但迄今为止，NDP52尚无明确的核功能。在这里，我们使用多学科方法来表征 NDP52 的生化特性和核作用。我们发现 NDP52 在转录起始位点与 RNA 聚合酶 II (RNAPII) 聚集，并且其过度表达促进其他转录簇的形成。我们还表明，NDP52 的缺失会影响两种模型哺乳动物细胞中的整体基因表达水平，并且转录抑制会影响细胞核中 NDP52 的空间组织和分子动力学。这直接将 NDP52 与 RNAPII 依赖性转录中的作用联系起来。此外，我们还表明 NDP52 与双链 DNA (dsDNA) 特异性且具有高亲和力结合，并且这种相互作用导致体外 DNA 结构的变化。这与我们的蛋白质组学数据（表明与核小体重塑蛋白和 DNA 结构调节剂相互作用的富集）一起表明 NDP52 在染色质调节中的可能功能。总的来说，我们在这里发现了 NDP52 在基因表达和 DNA 结构调控中的新核作用。
NDP52 is an autophagy receptor involved in the recognition and degradation of invading pathogens and damaged organelles. Although NDP52 was first identified in the nucleus and is expressed throughout the cell, to date, there is no clear nuclear function for NDP52. Here, we use a multidisciplinary approach to characterise the biochemical properties and nuclear roles of NDP52. We found that NDP52 clusters with RNA Polymerase II (RNAPII) at transcription initiation sites and that its overexpression promotes the formation of additional transcriptional clusters. We also show that depletion of NDP52 impacts overall gene-expression levels in two model mammalian cells, and that transcription inhibition affects the spatial organisation and molecular dynamics of NDP52 in the nucleus. This directly links NDP52 to a role in RNAPII-dependent transcription. Furthermore, we also show that NDP52 binds specifically and with high affinity to double-stranded DNA (dsDNA) and that this interaction leads to changes in DNA structure in vitro. This, together with our proteomics data indicating enrichment for interactions with nucleosome remodelling proteins and DNA structure regulators, suggests a possible function for NDP52 in chromatin regulation. Overall, here we uncover novel nuclear roles for NDP52 in gene expression and DNA structure regulation.