CLK-dependent exon recognition and conjoined gene formation revealed with a novel small molecule inhibitor
| Autor: | Damian Yap, Nao Morishita, Atsushi Nakanishi, Karey Shumansky, Shinsuke Araki, Shoichi Nakao, Satoshi Sasaki, Esther Kong, Ryujiro Hara, Momoko Ohori, Hiroyoshi Toyoshiba, Adrian Wan, Masatoshi Karashima, Mari Ogasawara-Shimizu, Osamu Nakanishi, Hirokazu Tozaki, Christopher S. Hughes, Noriko Uchiyama, S.-W. Grace Cheng, Masaya Sano, Daisuke Morishita, Tomohiro Kawamoto, Tohru Miyazaki, Shinya Tasaki, Jamie Rosner, Tyler Funnell, Sohrab P. Shah, Arusha Oloumi, Toshiyuki Nomura, Samuel Aparicio, Steven McKinney, Yusuke Nakayama, Tomohiro Ohashi, Aiko Murai, Gregg B. Morin, Misa Iwatani |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Transcription Genetic Science Exonic splicing enhancer General Physics and Astronomy 02 engineering and technology Protein Serine-Threonine Kinases Biology Article General Biochemistry Genetics and Molecular Biology Structure-Activity Relationship 03 medical and health sciences Exon Humans RNA Messenger Phosphorylation RNA Small Interfering Protein Kinase Inhibitors Gene Genetics Multidisciplinary Genome Human Kinase Gene Expression Profiling Alternative splicing Imidazoles Intron RNA-Binding Proteins Exons General Chemistry Protein-Tyrosine Kinases HCT116 Cells 021001 nanoscience & nanotechnology 3. Good health Alternative Splicing Pyrimidines 030104 developmental biology RNA splicing 0210 nano-technology |
| Zdroj: | Nature Communications Nature Communications, Vol 8, Iss 1, Pp 1-16 (2017) |
| ISSN: | 2041-1723 |
| Popis: | CDC-like kinase phosphorylation of serine/arginine-rich proteins is central to RNA splicing reactions. Yet, the genomic network of CDC-like kinase-dependent RNA processing events remains poorly defined. Here, we explore the connectivity of genomic CDC-like kinase splicing functions by applying graduated, short-exposure, pharmacological CDC-like kinase inhibition using a novel small molecule (T3) with very high potency, selectivity, and cell-based stability. Using RNA-Seq, we define CDC-like kinase-responsive alternative splicing events, the large majority of which monotonically increase or decrease with increasing CDC-like kinase inhibition. We show that distinct RNA-binding motifs are associated with T3 response in skipped exons. Unexpectedly, we observe dose-dependent conjoined gene transcription, which is associated with motif enrichment in the last and second exons of upstream and downstream partners, respectively. siRNA knockdown of CLK2-associated genes significantly increases conjoined gene formation. Collectively, our results reveal an unexpected role for CDC-like kinase in conjoined gene formation, via regulation of 3′-end processing and associated splicing factors. The phosphorylation of serine/arginine-rich proteins by CDC-like kinase is a central regulatory mechanism for RNA splicing reactions. Here, the authors synthesize a novel small molecule CLK inhibitor and map CLK-responsive alternative splicing events and discover an effect on conjoined gene transcription. |
| Databáze: | OpenAIRE |
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