This proposal seeks competitive renewal of a multi-PI project (Fu and Yeo), which aims to use global approaches to elucidate the regulatory principles of RNA binding proteins (RBPs) in mammalian genomes. Built upon our accomplishments in the past funding cycle, we propose to leverage the powerful experimental and computational tools we have developed to pursue four specific aims.
In Aim 1, we will couple gain- and lost-of-function multi-target screens to deduce regulatory pathways at both splicing and polyadenylation levels. We will focus on determining the specific function of different RNA polymerase II (Pol II) subunits, rather than by the Pol II CTD alone, in the recruitment of RNA processing machineries for co-transcriptional RNA processing.
In Aim 2, we will develop a general strategy for systematic identification of chromatin-associated RBPs to determine direct contribution of some RBPs to transcription and co-transcriptional RNA processing reactions. We will concentrate our efforts in dissecting a potential new pathway in epigenetic control of alternative splicing as well as broader roles of specific RBPs in direct transcriptional control.
In Aim 3, we will use a novel strategy for identification and characterization of non-canonical RBPs. Focusing on a large number of newly identified finger zinc (Znf) proteins, we propose to determine their roles in binding to both DNA and RNA and deduce their transcriptome-wide interactions with RNA. We also propose to pursue a specific paradigm in this aim on a newly identified RBP known to associate with the nuclear pore to determine its role in selective mRNA nuclear export, which is pertinent to an ALS-regulated disease pathology. Combined, we believe that this comprehensive, interconnected, and hypothesis-driven research plan will greatly advance our understanding of regulated RNA processing and associated disease mechanisms.
Our RNA genomics project aims to use genomics tools to systematically elucidate genes, gene networks, and pathways involved in the regulation of RNA processing in mammalian cells. The proposed research will provide critical molecular insights into regulated RNA processing and its coupling with other steps in gene expression, which will form the basis for development of effective treatment strategies against many RNA-related human diseases.
|Chen, Liang; Chen, Jia-Yu; Huang, Yi-Jou et al. (2018) The Augmented R-Loop Is a Unifying Mechanism for Myelodysplastic Syndromes Induced by High-Risk Splicing Factor Mutations. Mol Cell 69:412-425.e6|
|Markmiller, Sebastian; Soltanieh, Sahar; Server, Kari L et al. (2018) Context-Dependent and Disease-Specific Diversity in Protein Interactions within Stress Granules. Cell 172:590-604.e13|
|Bao, Xichen; Guo, Xiangpeng; Yin, Menghui et al. (2018) Capturing the interactome of newly transcribed RNA. Nat Methods 15:213-220|
|Nussbacher, Julia K; Yeo, Gene W (2018) Systematic Discovery of RNA Binding Proteins that Regulate MicroRNA Levels. Mol Cell 69:1005-1016.e7|
|Hu, Jing; Qian, Hao; Xue, Yuanchao et al. (2018) PTB/nPTB: master regulators of neuronal fate in mammals. Biophys Rep 4:204-214|
|Roloff, Alexander; Nelles, David A; Thompson, Matthew P et al. (2018) Self-Transfecting Micellar RNA: Modulating Nanoparticle Cell Interactions via High Density Display of Small Molecule Ligands on Micelle Coronas. Bioconjug Chem 29:126-135|
|Zhang, Kai; Zhang, Xiaorong; Cai, Zhiqiang et al. (2018) A novel class of microRNA-recognition elements that function only within open reading frames. Nat Struct Mol Biol 25:1019-1027|
|Wheeler, Emily C; Van Nostrand, Eric L; Yeo, Gene W (2018) Advances and challenges in the detection of transcriptome-wide protein-RNA interactions. Wiley Interdiscip Rev RNA 9:|
|Krach, Florian; Batra, Ranjan; Wheeler, Emily C et al. (2018) Transcriptome-pathology correlation identifies interplay between TDP-43 and the expression of its kinase CK1E in sporadic ALS. Acta Neuropathol 136:405-423|
|Van Nostrand, Eric L; Shishkin, Alexander A; Pratt, Gabriel A et al. (2017) Variation in single-nucleotide sensitivity of eCLIP derived from reverse transcription conditions. Methods 126:29-37|
Showing the most recent 10 out of 110 publications