JMJD6 cleaves MePCE to release positive transcription elongation factor b (P-TEFb) in higher eukaryotes
| Autor: | Xinjian Liu, Gongyi Zhang, Zhongzhou Chen, Frances Crawford, Chuan-Yuan Li, Haolin Liu, John W. Kappler, Philippa Marrack, Yuhua Xue, Schuyler Lee, Kathrin Maria Bernt, Qianqian Zhang, Chunjing Chen, Molly C. Kingsley, Xia Hong, Kirk C. Hansen, Ryan C. Hill, Qiang Zhou, Andreas Lengeling |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Mouse QH301-705.5 viruses Science Blotting Western Receptors Cell Surface RNA polymerase II CDK9 MePCE environment and public health Mass Spectrometry General Biochemistry Genetics and Molecular Biology P-TEFb Gene Knockout Techniques Mice 03 medical and health sciences 0302 clinical medicine Biochemistry and Chemical Biology Capping enzyme 7SK RNA JMJD6 Animals Positive Transcriptional Elongation Factor B snRNP Biology (General) 7SK snRNP Binding Sites General Immunology and Microbiology biology Chemistry General Neuroscience Methyltransferases General Medicine Protein Structure Tertiary 3. Good health Cell biology Bromodomain 030104 developmental biology 030220 oncology & carcinogenesis biology.protein Phosphorylation Medicine Cyclin-dependent kinase 9 Research Article |
| Zdroj: | eLife, Vol 9 (2020) eLife |
| Popis: | More than 30% of genes in higher eukaryotes are regulated by promoter-proximal pausing of RNA polymerase II (Pol II). Phosphorylation of Pol II CTD by positive transcription elongation factor b (P-TEFb) is a necessary precursor event that enables productive transcription elongation. The exact mechanism on how the sequestered P-TEFb is released from the 7SK snRNP complex and recruited to Pol II CTD remains unknown. In this report, we utilize mouse and human models to reveal methylphosphate capping enzyme (MePCE), a core component of the 7SK snRNP complex, as the cognate substrate for Jumonji domain-containing 6 (JMJD6)’s novel proteolytic function. Our evidences consist of a crystal structure of JMJD6 bound to methyl-arginine, enzymatic assays of JMJD6 cleaving MePCE in vivo and in vitro, binding assays, and downstream effects of Jmjd6 knockout and overexpression on Pol II CTD phosphorylation. We propose that JMJD6 assists bromodomain containing 4 (BRD4) to recruit P-TEFb to Pol II CTD by disrupting the 7SK snRNP complex. eLife digest In animals, an enzyme known as RNA polymerase II (Pol II for short) is a key element of the transcription process, whereby the genetic information contained in DNA is turned into messenger RNA molecules in the cells, which can then be translated to proteins. To perform this task, Pol II needs to be activated by a complex of proteins called P-TEFb; however, P-TEFb is usually found in an inactive form held by another group of proteins. Yet, it is unclear how P-TEFb is released and allowed to activate Pol II. Scientists have speculated that another protein called JMJD6 (Jumonji domain-containing 6) is important for P-TEFb to activate Pol II. Various roles for JMJD6 have been proposed, but its exact purpose remains unclear. Recently, two enzymes closely related to JMJD6 were found to be able to make precise cuts in other proteins; Lee, Liu et al. therefore wanted to test whether this is also true of JMJD6. Experiments using purified JMJD6 showed that it could make a cut in an enzyme called MePCE, which belongs to the group of proteins that hold P-TEFb in its inactive form. Lee, Liu et al. then tested the relationships between these proteins in living human and mouse cells. The levels of activated Pol II were lower in cells without JMJD6 and higher in those without MePCE. Together, the results suggest that JMJD6 cuts MePCE to release P-TEFb, which then activates Pol II. JMJD6 appears to know where to cut by following a specific pattern of elements in the structure of MePCE. When MePCE was mutated so that the pattern changed, JMJD6 was unable to cut it. These results suggest that JMJD6 and related enzymes belong to a new family of proteases, the molecular scissors that can cleave other proteins. The molecules that regulate transcription often are major drug targets, for example in the fight against cancer. Ultimately, understanding the role of JMJD6 might help to identify new avenues for cancer drug development. |
| Databáze: | OpenAIRE |
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