How does mRNA nucleoside modification impact protein production?
The modification of mRNAs has the potential to modulate every step in the mRNA life-cycle after transcription. The goal of our work is to understand how the molecular level details of mRNA modifications alter protein synthesis. To accomplish this, we use a combination of fully-purified translation assays together with lysate- and cell-based approaches to study how protein synthesis rates, translational fidelity, and mRNA stability are impacted by mRNA modifications.
Nucleoside modification discovery and mapping.
The current process to identify mRNA modifications is laborious; researchers hypothesize the presence of a particular modification and develop the antibody- and reverse transcriptase-based tools to map the position of the modification to the entire transcriptome. While this powerful approach has yielded ground-breaking findings is not a tractable way to quantitatively explore the full possible mRNA modification chemical space. We are developing mass-spectrometry and aptamer-based approaches for studying mRNA post-transcriptional modifications.
Collaborators: Dr. Qishan Lin (RNA Institute at SUNY Albany), Prof. Yingfu Li, McMaster University
How are modified nucleosides enzymatically incorporated?
We are using a combination of structural biology and mechanistic enzymology to study how mRNA modifying enzymes specifically select and modify their substrates. Our studies will provide the molecular level framework necessary for understanding how chemical modifications are inserted in protein coding mRNAs.
Collaborator: Prof. Markos Koutmos (University of Michigan)
What are the molecular level determinants of protein synthesis rates?
Despite being one of the most essential processes in biology there are still large gaps in our knowledge of how translation is regulated. We are using reconstituted translation systems and ribosome profiling to better understand how mRNA sequence, nascent peptide identity, and post-translational modification of the protein synthesis machinery impact translation. These studies will paint a broad molecular level picture of how translation elongation is altered to control gene expression.
Collaborators: Prof. Christopher Schofield (Oxford), Prof. Pavel Nagorny (University of Michigan)