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Abstract
The RNA guanine-N7 methyltransferase (RNMT) in complex with RNMT-Activating Miniprotein (RAM) catalyses the formation of a N7-methylated guanosine cap structure on the 5’ end of nascent RNA polymerase II transcripts. The mRNA cap protects the primary transcript from exonucleases and recruits cap-binding complexes that mediate RNA processing, export and translation. By using microsecond standard and accelerated molecular dynamics simulations, we provide for the first time a detailed molecular mechanism of allosteric regulation of RNMT by RAM. We show that RAM selects the RNMT active site conformations that are optimal for binding of substrates (AdoMet and the cap), thus enhancing their affinity. Furthermore, our results strongly suggest the likely scenario in which the cap binding promotes the subsequent AdoMet binding, consistent with the previously suggested cooperative binding model. By employing the network community analyses, we revealed the underlying long-range allosteric networks and paths that are crucial for allosteric regulation by RAM. Our findings complement and explain previous experimental data on RNMT activity. Moreover, this study provides the most complete description of the cap and AdoMet binding poses and interactions within the enzyme’s active site. This information is critical for the drug discovery efforts that consider RNMT as a promising anti-cancer target.
Original language | English |
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Article number | gkz613 |
Pages (from-to) | 8675-8692 |
Number of pages | 18 |
Journal | Nucleic Acids Research |
Volume | 47 |
Issue number | 16 |
DOIs | |
Publication status | Published - 19 Sept 2019 |
Keywords
- RNMT
- allosteric activation
- accelerated molecular dynamics
- conformational selection
- mRNA cap
- Allosteric Regulation
- Humans
- Substrate Specificity
- RNA Caps/chemistry
- Recombinant Proteins/chemistry
- Thermodynamics
- RNA Polymerase II/genetics
- Methyltransferases/chemistry
- S-Adenosylmethionine/chemistry
- Cloning, Molecular
- Transcription, Genetic
- Protein Interaction Domains and Motifs
- Binding Sites
- Genetic Vectors/chemistry
- Amino Acid Sequence
- Protein Conformation, alpha-Helical
- Gene Expression
- RNA-Binding Proteins/chemistry
- S-Adenosylhomocysteine/chemistry
- Escherichia coli/genetics
- Molecular Dynamics Simulation
- Sequence Homology, Amino Acid
- Sequence Alignment
- Protein Conformation, beta-Strand
- Protein Binding
- Kinetics
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Dive into the research topics of 'Mechanism of allosteric activation of human mRNA cap methyltransferase (RNMT) by RAM: Insights from accelerated molecular dynamics simulations'. Together they form a unique fingerprint.Projects
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Student theses
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Characterising the recruitment and activation mechanisms of the mRNA capping enzymes via molecular dynamics simulation
Author: Bage, M. G., 2021Supervisor: Pisliakov, A. (Supervisor) & Cowling, V. (Supervisor)
Student thesis: Doctoral Thesis › Doctor of Philosophy
Profiles
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Cowling, Victoria
- Molecular Cell and Developmental Biology - Associate Staff of Biology
Person: Associate Staff