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M6a dot blot protocol12/1/2023 Circadian control of the NAD + salvage pathway by CLOCK-SIRT1. Nakahata, Y., Sahar, S., Astarita, G., Kaluzova, M. Extraction and quantitation of nicotinamide adenine dinucleotide redox cofactors. NAD(H) and NADP(H) redox couples and cellular energy metabolism. The obesity-associated FTO gene encodes a 2-oxoglutarate-dependent nucleic acid demethylase. FTO-mediated formation of N 6-hydroxymethyladenosine and N 6-formyladenosine in mammalian RNA. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. N 6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Internal mRNA methylation finally finds functions. Where, when and how: context-dependent functions of RNA methylation writers, readers and erasers. Topology of the human and mouse m 6A RNA methylomes revealed by m 6A-seq. Genome-wide association studies provide new insights into type 2 diabetes aetiology. Genome-wide association yields new sequence variants at seven loci that associate with measures of obesity. Ribozymes, riboswitches and beyond: regulation of gene expression without proteins. Hypoxia-inducible factors in physiology and medicine. How cancer metabolism is tuned for proliferation and vulnerable to disruption. Cellular metabolism and induced pluripotency. These results build a direct link between metabolism and RNA m 6A demethylation. Furthermore, NADP regulated mRNA m 6A via FTO in vivo, and deletion of FTO blocked NADP-enhanced adipogenesis in 3T3-L1 preadipocytes. In vitro demethylation assays indicated that NADP enhances FTO activity. We screened a set of metabolites using a fluorescence quenching assay and NADP was identified to remarkably bind FTO. Here, we show that NADP directly binds FTO, independently increases FTO activity, and promotes RNA m 6A demethylation and adipogenesis. However, knowledge about whether and which metabolites directly regulate m 6A remains elusive. This hypothesis is at least partially supported by the findings that multiple metabolic diseases are highly associated with fat mass and obesity-associated protein (FTO), an m 6A demethylase. In turn, it is likely that some metabolites regulate enzymes controlling reversible RNA modification, such as N 6-methyladenosine (m 6A), to modulate RNA. Metabolism is often regulated by the transcription and translation of RNA.
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