Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5589907 | Gene | 2017 | 18 Pages |
Abstract
Gene regulation at the post-transcriptional level is frequently based on cis- and trans-acting factors on target mRNAs. We found a C-rich element (CRE) in mu-opioid receptor (MOR) 3â²-untranslated region (UTR) to which poly (rC) binding protein 1 (PCBP1) binds, resulting in MOR mRNA stabilization. RNA immunoprecipitation and RNA EMSA revealed the formation of PCBP1-RNA complexes at the element. Knockdown of PCBP1 decreased MOR mRNA half-life and protein expression. Stimulation by forskolin increased cytoplasmic localization of PCBP1 and PCBP1/MOR 3â²-UTR interactions via increased serine phosphorylation that was blocked by protein kinase A (PKA) or (phosphatidyl inositol-3) PI3-kinase inhibitors. The forskolin treatment also enhanced serine- and tyrosine-phosphorylation of AU-rich element binding protein (AUF1), concurrent with its increased binding to the CRE, and led to an increased interaction of poly A binding protein (PABP) with the CRE and poly(A) sites. AUF1 phosphorylation also led to an increased interaction with PCBP1. These findings suggest that a single co-regulator, PCBP1, plays a crucial role in stabilizing MOR mRNA, and is induced by PKA signaling by conforming to AUF1 and PABP.
Keywords
PCBP1DAB2LMBIRESphorbol 12-myristate 13-acetateRNPsPABPRibonucleoproteinsPKCrpl32CREpKahuman antigen RAUF1LUCSTRMOR3′-UTR3′-untranslated regionhnRNPsPMAsodium dodecyl sulfate-polyacrylamide gel electrophoresisSDS-PAGELOXRNA immunoprecipitationDiCEShort tandem repeatsRNA stabilityHeterogeneous nuclear ribonucleoproteinsAU-rich elementsluciferaseLeptomycin BlipoxygenaseDisabled-2NucleotideARERIPribosomal protein L32RNA binding proteinprotein kinase AProtein kinase CHuRMu opioid receptor
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Authors
Cheol Kyu Hwang, Yadav Wagley, Ping-Yee Law, Li-Na Wei, Horace H. Loh,