4 research outputs found
Proceedings of the Thirteenth International Society of Sports Nutrition (ISSN) Conference and Expo
Meeting Abstracts: Proceedings of the Thirteenth International Society of Sports Nutrition (ISSN) Conference and Expo Clearwater Beach, FL, USA. 9-11 June 201
Lysines in the RNA Polymerase II C鈥慣erminal Domain Contribute to TAF15 Fibril Recruitment
Many cancer-causing chromosomal translocations
result in transactivating
protein products encoding FET family (FUS, EWSR1, TAF15) low-complexity
(LC) domains fused to a DNA binding domain from one of several transcription
factors. Recent work demonstrates that higher-order assemblies of
FET LC domains bind the carboxy-terminal domain of the large subunit
of RNA polymerase II (RNA pol II CTD), suggesting FET oncoproteins
may mediate aberrant transcriptional activation by recruiting RNA
polymerase II to promoters of target genes. Here we use nuclear magnetic
resonance (NMR) spectroscopy and hydrogel fluorescence microscopy
localization and fluorescence recovery after photobleaching to visualize
atomic details of a model of this process, interactions of RNA pol
II CTD with high-molecular weight TAF15 LC assemblies. We report NMR
resonance assignments of the intact degenerate repeat half of human
RNA pol II CTD alone and verify its predominant intrinsic disorder
by molecular simulation. By measuring NMR spin relaxation and dark-state
exchange saturation transfer, we characterize the interaction of RNA
pol II CTD with amyloid-like hydrogel fibrils of TAF15 and hnRNP A2
LC domains and observe that heptads far from the acidic C-terminal
tail of RNA pol II CTD bind TAF15 fibrils most avidly. Mutation of
CTD lysines in heptad position 7 to consensus serines reduced the
overall level of TAF15 fibril binding, suggesting that electrostatic
interactions contribute to complex formation. Conversely, mutations
of position 7 asparagine residues and truncation of the acidic tail
had little effect. Thus, weak, multivalent interactions between TAF15
fibrils and heptads throughout RNA pol II CTD collectively mediate
complex formation
Recommended from our members
A guide to the BRAIN Initiative Cell Census Network data ecosystem
Characterizing cellular diversity at different levels of biological organization and across data modalities is a prerequisite to understanding the function of cell types in the brain. Classification of neurons is also essential to manipulate cell types in controlled ways and to understand their variation and vulnerability in brain disorders. The BRAIN Initiative Cell Census Network (BICCN) is an integrated network of data-generating centers, data archives, and data standards developers, with the goal of systematic multimodal brain cell type profiling and characterization. Emphasis of the BICCN is on the whole mouse brain with demonstration of prototype feasibility for human and nonhuman primate (NHP) brains. Here, we provide a guide to the cellular and spatial approaches employed by the BICCN, and to accessing and using these data and extensive resources, including the BRAIN Cell Data Center (BCDC), which serves to manage and integrate data across the ecosystem. We illustrate the power of the BICCN data ecosystem through vignettes highlighting several BICCN analysis and visualization tools. Finally, we present emerging standards that have been developed or adopted toward Findable, Accessible, Interoperable, and Reusable (FAIR) neuroscience. The combined BICCN ecosystem provides a comprehensive resource for the exploration and analysis of cell types in the brain