Mapping the micro-proteome of the nuclear lamina and lamina-associated domains

Autor: Jevon Cutler, Xianrong Wong, Karen L. Reddy, Anil K. Madugundu, Molly Gordon, Akhilesh Pandey, Victoria E. Hoskins
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Life Science Alliance
ISSN: 2575-1077
Popis: 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.
Databáze: OpenAIRE