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

OBJECTIVE: Glucagon in mammals and its homolog (adipokinetic hormone [AKH] in Drosophila melanogaster) 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 gen...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society of Lipidology and Atherosclerosis 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792818/
https://www.ncbi.nlm.nih.gov/pubmed/35118022
http://dx.doi.org/10.12997/jla.2022.11.1.55
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author 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
author_facet 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
author_sort Ko, Seungbeom
collection PubMed
description OBJECTIVE: Glucagon in mammals and its homolog (adipokinetic hormone [AKH] in Drosophila melanogaster) 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 Gcg messenger RNA (mRNA) in mouse and Akh mRNA in Drosophila. METHODS: Drosophila Schneider 2 (S2) and mouse 3T3-L1 cell lysates were utilized for affinity pull down of Akh and Gcg 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. RESULTS: We observed that 1) 91 proteins interact with Akh mRNA from S2 cell lysates, 2) 34 proteins interact with Gcg mRNA from 3T3-L1 cell lysates. 3) Akh mRNA interactome revealed clusters of ribosomes and known RNA-binding proteins (RBPs). 4) Gcg mRNA interactome revealed mRNA-binding proteins including Plekha7, zinc finger protein, carboxylase, lipase, histone proteins and a cytochrome, Cyp2c44. 5) Levels of Gcg mRNA and its interacting proteins are elevated in skeletal muscles isolated from old mice compared to ones from young mice. CONCLUSION: Akh mRNA in S2 cells are under active translation in a complex of RBPs and ribosomes. Gcg mRNA in mouse precursor adipocyte is in a condition distinct from Akh mRNA due to biochemical interactions with a subset of RBPs and histones. We anticipate that our study contributes to investigating regulatory mechanisms of Gcg and Akh mRNA decay, translation, and localization.
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spelling pubmed-87928182022-02-02 Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs 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 J Lipid Atheroscler Original Article OBJECTIVE: Glucagon in mammals and its homolog (adipokinetic hormone [AKH] in Drosophila melanogaster) 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 Gcg messenger RNA (mRNA) in mouse and Akh mRNA in Drosophila. METHODS: Drosophila Schneider 2 (S2) and mouse 3T3-L1 cell lysates were utilized for affinity pull down of Akh and Gcg 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. RESULTS: We observed that 1) 91 proteins interact with Akh mRNA from S2 cell lysates, 2) 34 proteins interact with Gcg mRNA from 3T3-L1 cell lysates. 3) Akh mRNA interactome revealed clusters of ribosomes and known RNA-binding proteins (RBPs). 4) Gcg mRNA interactome revealed mRNA-binding proteins including Plekha7, zinc finger protein, carboxylase, lipase, histone proteins and a cytochrome, Cyp2c44. 5) Levels of Gcg mRNA and its interacting proteins are elevated in skeletal muscles isolated from old mice compared to ones from young mice. CONCLUSION: Akh mRNA in S2 cells are under active translation in a complex of RBPs and ribosomes. Gcg mRNA in mouse precursor adipocyte is in a condition distinct from Akh mRNA due to biochemical interactions with a subset of RBPs and histones. We anticipate that our study contributes to investigating regulatory mechanisms of Gcg and Akh mRNA decay, translation, and localization. Korean Society of Lipidology and Atherosclerosis 2022-01 2021-07-29 /pmc/articles/PMC8792818/ /pubmed/35118022 http://dx.doi.org/10.12997/jla.2022.11.1.55 Text en Copyright © 2022 The Korean Society of Lipid and Atherosclerosis. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
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
Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title_full Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title_fullStr Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title_full_unstemmed Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title_short Profiling of RNA-binding Proteins Interacting With Glucagon and Adipokinetic Hormone mRNAs
title_sort profiling of rna-binding proteins interacting with glucagon and adipokinetic hormone mrnas
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8792818/
https://www.ncbi.nlm.nih.gov/pubmed/35118022
http://dx.doi.org/10.12997/jla.2022.11.1.55
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