Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs

• Published in: Journal of lipid and atherosclerosis Vol. 11; no. 1; pp. 55 - 72
• Main Authors: Ko, Seungbeom, Yeom, Eunbyul, Chun, Yoo Lim, Mun, Hyejin, Howard-McGuire, Marina, Millison, Nathan T, Jung, Junyang, Lee, Kwang-Pyo, Lee, Changhan, Lee, Kyu-Sun, Delaney, Joe R, Yoon, Je-Hyun
• Format: Journal Article
• Language: English
• Published: Korea (South) Korean Society of Lipidology and Atherosclerosis 01.01.2022; 한국지질동맥경화학회
• Subjects: Original; 내과학; Glucagon; Mass spectrometry; RNA-binding proteins
• ISSN: 2287-2892; 2288-2561
• DOI: 10.12997/jla.2022.11.1.55

Glucagon in mammals and its homolog (adipokinetic hormone [AKH] in ) are peptide hormones which regulate lipid metabolism by breaking down triglycerides. Although regulatory mechanisms of glucagon and AKH expression have been widely studied, post-transcriptional gene expression of glucagon has not been investigated thoroughly. In this study, we aimed to profile proteins binding with messenger RNA (mRNA) in mouse and mRNA in Drosophila. Drosophila Schneider 2 (S2) and mouse 3T3-L1 cell lysates were utilized for affinity pull down of and mRNA respectively using biotinylated anti-sense DNA oligoes against target mRNAs. Mass spectrometry and computational network analysis revealed mRNA-interacting proteins residing in functional proximity. We observed that 1) 91 proteins interact with mRNA from S2 cell lysates, 2) 34 proteins interact with mRNA from 3T3-L1 cell lysates. 3) mRNA interactome revealed clusters of ribosomes and known RNA-binding proteins (RBPs). 4) mRNA interactome revealed mRNA-binding proteins including Plekha7, zinc finger protein, carboxylase, lipase, histone proteins and a cytochrome, Cyp2c44. 5) Levels of mRNA and its interacting proteins are elevated in skeletal muscles isolated from old mice compared to ones from young mice. mRNA in S2 cells are under active translation in a complex of RBPs and ribosomes. mRNA in mouse precursor adipocyte is in a condition distinct from mRNA due to biochemical interactions with a subset of RBPs and histones. We anticipate that our study contributes to investigating regulatory mechanisms of and mRNA decay, translation, and localization.