Abstract

CCAAT/enhancer-binding protein beta (C/EBPβ) is a transcription factor controlling a broad range of genes essential for homeostasis, including genes related to immune functions, inflammation, metabolism and growth. Monocyte chemoattractant protein-1-induced protein 1 (MCPIP1) also called as Regnase-1 is an RNase and has been shown to decrease the stability of short-lived transcripts coding for inflammation-related proteins, including IL-1β, IL-6, IL-2, IL-8, IL-12b, IER-3, c-Rel. We found previously that the half-life of the C/EBPβ transcript is regulated by MCPIP. To understand the mechanism driving down-regulation of C/EBPβ by MCPIP1, we applied an in vitro cleavage assay, followed by a luciferase-reporter assay and RNA immunoprecipitation (RIP). We demonstrated that MCPIP1 recognizes regions of the 3’UTR of C/EBPβ mRNA and promotes its decay by introducing direct endonucleolytic cleavage.

Highlights

  • Modulation of mRNA stability has been found to be responsible for 40–50% of all changes in gene expression [1,2]

  • To specify which 3’ untranslated region of an mRNA (3’UTR) region of the C/EBPβ mRNA is involved in the Monocyte chemoattractant protein-1-induced protein 1 (MCPIP1)-driven down-regulation, we prepared a set of reporter vectors harboring different sections of the 3’UTR of C/EBPβ mRNA (Fig 1A) which were linked to a luciferase coding sequence (CDS)

  • Constructs were introduced to HepG2 cells together with plasmids coding for MCPIP1 or MCPIP1 lacking RNase activity (MCPIP1-D141N)

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Summary

Introduction

Modulation of mRNA stability has been found to be responsible for 40–50% of all changes in gene expression [1,2] Research in this field has revealed the importance of cis-acting elements located predominantly in the 3’ untranslated region of an mRNA (3’UTR). These elements enable the interaction of mRNA transcripts with an mRNA-decay complex where unspecific nucleases promote degradation. A recently discovered endonuclease capable of mRNA cleavage is known as MCPIP1. This protein is essential for the degradation of short-lived transcripts coding for inflammation-related proteins, including IL-1β, IL-6, IL-2, IL-8, IL-12b, IER-3, c-Rel [4,5,6,7,8].

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