Biology subjects

Wei, F.-Y.

Publications and source records attributed to Wei, F.-Y..

6 recordsLinked to original sources

bioRxiv2024

A sequential, RNA-derived, modified adenosine pathway safeguards cellular metabolism

RNA contains diverse post-transcriptional modifications and its catabolic breakdown yields numerous modified nucleosides that must be properly processed, but the molecular mechanism is largely unknown. Here, we show that three RNA-derived modified adenosines, N6-methyladenosine (m6A), N6,N6-dimethyladenosine (m6,6A), and N6-isopentenyladenosine (i6A), are sequentially metabolized to inosine monophosphate (IMP) to prevent their intrinsic cytotoxicity. These modified adenosines are phosphorylated by adenosine kinase (ADK) followed by adenosine deaminase-like (ADAL)-mediated deamination in both plants and animals. ADAL knockout mice accumulate N6-modified AMPs that allosterically inhibit AMP-activated protein kinase (AMPK), leading to dysregulation of glucose metabolism. Furthermore, ADK deficiency, reported in patients with severe metabolic defects, induces aberrant elevation of m6A/m6,6A/i6A, disrupting lipid metabolism and causing early death in mouse models. The findings unveil a fundamental mechanism by which cells alleviate the toxicity of modified adenosines, and that connects modified adenosines to human disease. In briefRNA catabolism yields diverse nucleosides with modifications attached. Three RNA-derived modified adenosines (m6A, m6,6A, and i6A) are intrinsically toxic and subjected to sequential metabolism to yield inosine monophosphate. Dysregulation of this pathway impairs energy balance and contributes to metabolic diseases. HighlightsO_LICytotoxic RNA catabolism-derived modified adenosines undergo sequential metabolism C_LIO_LIm6A, m6,6A, and i6A are phosphorylated by ADK then deaminated by ADAL to IMP C_LIO_LIm6AMP, m6,6AMP, and i6AMP allosterically inhibit AMP-mediated AMPK activation C_LIO_LILoss of ADK causes the accumulation of m6A, m6,6A, and i6A, and abnormal lipid metabolism, which can lead to genetic disease C_LI

cell biology↗

bioRxiv2024

Structural insights into the agonist selectivity and structure-based engineering of the adenosine A3 receptor

Adenosine receptors, expressed across various tissues, play pivotal roles in physiological processes and are implicated in diverse diseases, including neurological disorders and inflammation, highlighting the therapeutic potential of receptor-selective agents. The Adenosine A3 receptor (A3R), the last identified adenosine receptor, is also activated by breakdown products of post-transcriptionally modified tRNA and exhibits dual roles in neuron, heart, and immune cells, and is often overexpressed in tumors, making it a target for anticancer therapy. Despite extensive studies on the other adenosine receptors, the structure and activation mechanism of A3R, especially by selective agonists like N6-methyladenosine (m6A) and namodenoson, remained elusive. Here, we identified N6-isopentenyl adenosine (i6A), a novel A3R-selective ligand, via comprehensive modified adenosine library screening. Cryo-EM analyses of A3R-Gi signaling complexes with two nonselective and three selective agonists revealed the structural basis for A3R activation. We further conducted structure-guided engineering of m6A-insensitive A3R, which would greatly facilitate future discoveries of the physiological functions of the selective activation of A3R by modified adenosines. Our results clarify the selective activation of adenosine receptors, providing the basis for future drug discovery.

biophysics↗

bioRxiv2023

Complexity and dynamics of in organello translation landscape assessed by high-resolution mitochondrial ribosome profiling

Since mitochondrial translation serves the essential subunits of the OXPHOS complex that produces ATP, exhaustive, quantitative, and high-resolution delineation of mitoribosome traversal is needed. Here, we developed a technique for high-resolution mitochondrial ribosome profiling and revealed the intricate regulation of mammals in organello translation. Our approach assessed the stoichiometry and kinetics of mitochondrial translation flux, such as the number of mitoribosomes on a transcript and the elongation rate, initiation rate, and lifetime rounds of translation of individual transcripts. We also surveyed the impacts of modifications at the anticodon stem loop in mt-tRNAs, including all possible modifications at the 34th position, by deleting the corresponding enzymes and harnessing patient-derived cells. Moreover, a retapamulin-assisted derivative and mito-disome profiling revealed cryptic translation initiation sites at subcognate codons and programmed mitoribosome collision sites across the mitochondrial transcriptome. Our work provides a useful platform for investigating protein synthesis within the energy powerhouse of the cell.

molecular biology↗

bioRxiv2022

Activation of the urotensin-II receptor by anti-COVID-19 drug remdesivir induces cardiomyocyte dysfunction

Remdesivir is an antiviral drug used for COVID-19 treatment worldwide. Cardiovascular (CV) side effects have been associated with remdesivir; however, the underlying molecular mechanism remains unknown. Here, we performed a large-scale G-protein-coupled receptor (GPCR) screening in combination with structural modeling and found that remdesivir is a selective agonist for urotensin-II receptor (UTS2R). Functionally, remdesivir treatment induced prolonged field potential in human induced pluripotent stem cell (iPS)-derived cardiomyocytes and reduced contractility in neonatal rat cardiomyocytes, both of which mirror the clinical pathology. Importantly, remdesivir-mediated cardiac malfunctions were effectively attenuated by antagonizing UTS2R signaling. Finally, we characterized the effect of 110 single-nucleotide variants (SNVs) in UTS2R gene reported in genome database and found four missense variants that show gain-of-function effects in the receptor sensitivity to remdesivir. Collectively, our study illuminates a previously unknown mechanism underlying remdesivir-related CV events and that genetic variations of UTS2R gene can be a potential risk factor for CV events during remdesivir treatment, which collectively paves the way for a therapeutic opportunity to prevent such events in the future. One Sentence SummaryRemdesivir s activity as a selective agonist of urotensin-II receptor underlies its known cardiotoxicity in anti-viral therapy.

pharmacology and toxicology↗

bioRxiv2020

N6-methyladenosine (m6A) is an endogenous A3 adenosine receptor ligand

About 150 post-transcriptional RNA modifications have been identified in all kingdoms of life. During RNA catabolism, most modified nucleosides are resistant to degradation and are released into the extracellular space. In this study, we explored the physiological role of these extracellular modified nucleosides and found that N6-methyladenosine (m6A), widely known as an epigenetic mark in RNA, acts as a ligand for the adenosine A3 receptor, for which it has greater affinity than unmodified adenosine. Structural modeling defined the amino acids required for specific binding of m6A to the A3 receptor. m6A is dynamically released in response to cytotoxic stimuli and facilitates type I allergy. Our findings shed light on m6A as a signaling molecule with the ability to activate GPCRs, a previously unreported property of RNA modifications.

molecular biology↗

bioRxiv2019

Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs

Post-transcriptional modifications in mitochondrial tRNAs (mt-tRNAs) play critical roles in mitochondrial protein synthesis, which produces respiratory chain complexes. In this study, we used mass spectrometric analysis to map 5-methylcytidine (m5C) at positions 48-50 in eight mouse and six human mt-tRNAs. We also confirmed the absence of m5C in mt-tRNAs isolated from Nsun2 knockout (KO) mice, as well as from NSUN2 KO human culture cells. In addition, we successfully reconstituted m5C at positions 48-50 of mt-tRNA in vitro with NSUN2 protein in the presence of S-adenosylmethionine (SAM). Although NSUN2 is predominantly localized to the nucleus and introduces m5C into cytoplasmic tRNAs and mRNAs, structured illumination microscopy (SIM) clearly revealed NSUN2 foci inside mitochondria. These observations provide novel insights into the role of NSUN2 in the physiology and pathology of mitochondrial functions.

molecular biology↗