Virginia Research Day 2021

Graduate Student Research Biomedical

11 Influence of Folate on RNA Cytosine Methylation in Neural Stem Cells

Xiguang Xu 1,2 ; Zachary Johnson 1,3 ; Razan Alajoleen 1,4 ; Xiaoran Wei 1,4 ; Natalie Melville 1,5 ; Amanda Wang 1 ; Rachel Padget 2,6 ; James Smyth 2,6 ; Hrubec Terry 4,7 ; Hehuang Xie 1,2,3,4,5* Corresponding author: arazan@vt.edu

1 Epigenomics and Computational Biology Lab, Fralin Life Sciences Institute of Virginia Tech, 2 Department of Biological Sciences, College of Science, Virginia Tech, 3 Genetics, Bioinformatics and Computational Biology Program, Virginia Tech, 4 Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine; Virginia Tech 5 Translational Biology, Medicine and Health Program, Virginia Tech, 6 Fralin Biomedical Research Institute at VTC, Virginia Tech, 7 Via College of Osteopathic Medicine-Virginia Campus

The study of RNA cytosine methylation (5-mrC) belongs to an emerging field of epitranscriptomics. Recent studies indicated that 5-mrC occurs in brain mRNAs coding for genes involved in central nervous system development and axonogenesis. Folate is a methylation donor and the suboptimal intake of this micronutrient has been linked to neural tube defect at embryonic stage and autism in early childhood. We hypothesize that the intake of folate may have an impact on brain epitranscriptomic profiles during development. In this study, we generated transcriptome-wide landscapes of 5-mrC modification in both total mRNAs and polysome- associated mRNAs for mouse neural stem cells

(NSCs) cultured in media with different levels of folate. Distinct 5-mrC modification profiles in both total mRNAs and polysome mRNAs were detected in NSCs cultured with different concentrations of folate. Interestingly, 5-mrC was found to be significantly enriched in polysome mRNAs over total mRNAs; and panel of differentially translated mRNAs carrying hypermethylated 5-mrC was revealed by polysome profiling in combination of bisulfite sequencing. In summary, this study provides the first comprehensive set of epitranscriptomic data to evaluate RNA transcription, RNA methylation, and RNA translation together in NSCs with both low and high folate treatment.

This study was funded in part by the VCOM-VMCVM One Health Program.

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