A sweet regulatory landscape of the glycome

• Author / Creator

  Chu, Thu

• Glycosylation is one of the most abundant and diverse post-translational modifications. Glycans participate in various key biological functions and processes including immunity, cell adhesion, endocytosis, exocytosis, molecular trafficking, and signal transduction. Previous work from our lab identified microRNAs (miRNAs, miRs) as key regulators of glycosylation genes (glycogenes). miRs are small non-coding RNAs that fine-tune protein expression through binding to messenger RNA (mRNA). miRs regulate networks of genes that work to control a specific biological process, tightening the expression range for critical genes. By mapping the targets of miRNA involved in specific biological processes or disease states, we can determine genes that are important drivers in that process or disease, defined as the miRNA Proxy Approach 1, 2. However, this approach is dependent on an accurate understanding of miRNA:mRNA interactions.
  
  Current identification of miR:glycogene interactomes is hindered by the low accuracy of prediction (17-66%), the low expression of glycogenes (complicating transcriptomic analysis and RISC complex pulldowns), and suboptimal throughput ((only around 0.01% of all predicted human miR:target interactions have been validated experimentally) of more direct miR:mRNA validations (mostly luciferase assay). To overcome these obstacles, our laboratory has developed miRFluR high-throughput experimental mapping of miR:glycogene interactions. Thus, we were able to obtain a comprehensive dataset of miRNA regulatory networks for glycosylation enzymes including B3GLCT, OGT and OGA. In the work of miR-B3GLCT interaction network, we successfully utilized downregulatory miR network to predict B3GLCT biological functions as supporting evidence for our miRNA proxy hypothesis 3. In addition, we not only identified miR impacting in the 3’ untranslated region (3’UTR) but also expanding our platform to the 5’UTR. In summary, this work contributed to decipher the glycosylation code and understanding biological functions of certain regulatory networks.

• Subjects / Keywords

  • miRNA
  • mRNA
  • Interactomes
  • Glycosylation
  • Glycoenzymes
  • Sweet regulation
  • miRFluR

• Graduation date

  Spring 2023

• Type of Item

  Thesis

• Degree

  Doctor of Philosophy

• DOI

  https://doi.org/10.7939/r3-5xqb-vx73

• License

  This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.