Integrative genomics reveal a role for MCPIP1 in adipogenesis and adipocyte metabolism

Magdalena Losko,Natalia Pydyn,Sylwia Kedracka-Krok,Jolanta Jura,Andrzej Budzynski,Agnieszka Paziewska,Piotr Major,Urszula Jankowska,Maria Kulecka,Marek Winiarski,Dobrochna Dolicka,Michal Mikula

doi:10.1007/s00018-019-03434-5

Abstract

Obesity is considered a serious chronic disease, associated with an increased risk of developing cardiovascular diseases, non-alcoholic fatty liver disease and type 2 diabetes. Monocyte chemoattractant protein-1-induced protein-1 (MCPIP1) is an RNase decreasing stability of transcripts coding for inflammation-related proteins. In addition, MCPIP1 plays an important role in the regulation of adipogenesis in vitro by reducing the expression of key transcription factors, including C/EBPβ. To elucidate the role of MCPIP1 in adipocyte biology, we performed RNA-Seq and proteome analysis in 3T3-L1 adipocytes overexpressing wild-type (WTMCPIP1) and the mutant form of MCPIP1 protein (D141NMCPIP1). Our RNA-Seq analysis followed by confirmatory Q-RT-PCR revealed that elevated MCPIP1 levels in 3T3-L1 adipocytes upregulated transcripts encoding proteins involved in signal transmission and cellular remodeling and downregulated transcripts of factors involved in metabolism. These data are consistent with our proteomic analysis, which showed that MCPIP1 expressing adipocytes exhibit upregulation of proteins involved in cellular organization and movement and decreased levels of proteins involved in lipid and carbohydrate metabolism. Moreover, MCPIP1 adipocytes are characterized by decreased level of insulin receptor, reduced insulin-induced Akt phosphorylation, as well as depleted Glut4 level and impaired glucose uptake. Overexpression of Glut4 in 3T3-L1 cells expressed WTMCPIP1 rescued adipogenesis. Interestingly, we found decreased level of MCPIP1 along with an increase in body mass index in subcutaneous adipose tissue. The presented data show a novel role of MCPIP1 in modulating insulin sensitivity in adipocytes. Overall, our findings demonstrate that MCPIP1 is an important regulator of adipogenesis and adipocyte metabolism.

Highlights

Monocyte chemoattractant protein-1-induced protein-1 (MCPIP1), known as Regnase-1, is an RNase controlling stability of many transcripts and miRNAs
Our comparative analysis done for two studied reference control genes EF2 and ACTB, has revealed lower deviations in Ct values for EF2 in analyzed samples in comparison to Ct values for ACTB (Supplementary data, Fig. 1a), EF2 was used as an internal control for the estimation of mRNA levels in adipose tissue biopsies
We observed statistically significant increase in the MCPIP1 transcript levels in subcutaneous adipose tissue (SAT) of lean subjects in comparison to obese subjects

Summary

Introduction

Monocyte chemoattractant protein-1-induced protein-1 (MCPIP1), known as Regnase-1, is an RNase controlling stability of many transcripts and miRNAs. The ribonucleolytic activity of MCPIP1 is associated with the PIN domain—named after its identification in the N-terminus of the PilT protein (PilT N-terminus). The ribonucleolytic activity of MCPIP1 is associated with the PIN domain—named after its identification in the N-terminus of the PilT protein (PilT N-terminus) Another domain essential for effective cleavage of RNA molecules is the zinc finger domain. In the case of miRNA, MCPIP1 degrades precursor transcripts, effectively terminating microRNA biogenesis [3]. MCPIP1 regulates processes important in cancer development, such as cell metabolism, proliferation, and angiogenesis [8,9,10]

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Results

Conclusion