Translation control of TAK1 mRNA by hnRNP K modulates LPS-induced macrophage activation

Liepelt, Anke; Mossanen, Jana C.; Denecke, Bernd; Heymann, Felix; Santis, Rebecca De; Tacke, Frank; Marx, Gernot; Ostareck, Dirk H.; Ostareck-Lederer, Antje

doi:10.1261/rna.042788.113

Cited by 30 publications

(44 citation statements)

References 61 publications

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“…Moreover, depletion of AUF1 from a leukemic cell line (THP-1) (42) somewhat increases TAK1 levels and stimulates NF-B-mediated gene transcription. In addition, a recent report demonstrated that depletion of hnRNP K also increases TAK1 levels (43). Because these observations are similar to those obtained with CPEB KO mice and cultured cells lacking CPEB, we hypothesize that there might be a causal link between CPEB, AUF1, hnRNP K, and TAK1-dependent inflammation.…”

Section: Discussionsupporting

confidence: 85%

CPEB Regulation of TAK1 Synthesis Mediates Cytokine Production and the Inflammatory Immune Response

Ivshina

Alexandrov

Vertii

et al. 2015

Molecular and Cellular Biology

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The cytoplasmic-element-binding (CPEB) protein is a sequence-specific RNA-binding protein that regulates cytoplasmic polyadenylation-induced translation. In mouse embryo fibroblasts (MEFs) lacking CPEB, many mRNAs encoding proteins involved in inflammation are misregulated. Correlated with this aberrant translation in MEFs, a macrophage cell line depleted of CPEB and treated with lipopolysaccharide (LPS) to stimulate the inflammatory immune response expresses high levels of interleukin-6 (IL-6), which is due to prolonged nuclear retention of NF-B. Two proteins involved in NF-B nuclear localization and IL-6 expression, IB␣ and transforming growth factor beta-activated kinase 1 (TAK1), are present at excessively low and high steadystate levels, respectively, in LPS-treated CPEB-depleted macrophages. However, only TAK1 has an altered synthesis rate that is CPEB dependent and CPEB/TAK1 double depletion alleviates high IL-6 production. Peritoneal macrophages isolated from CPEB knockout (KO) mice treated with LPS in vitro also have prolonged NF-B nuclear retention and produce high IL-6 levels. LPS-injected CPEB KO mice secrete prodigious amounts of IL-6 and other proinflammatory cytokines and exhibit hypersensitivity to endotoxic shock; these effects are mitigated when the animals are also injected with (5Z)-7-oxozeaenol, a potent and specific inhibitor of TAK1. These data show that CPEB control of TAK1 mRNA translation mediates the inflammatory immune response. Inflammation is triggered by bacterial pathogens, as well as lipopolysaccharide (LPS), a bacterial cell wall component that activates the transcription of multiple inflammatory response genes, including those for cytokines and chemokines (1, 2). Despite the importance of these inflammation mediators for host defense against infection, their excessive production can elicit organ failure and septic shock that results in lethality. Therefore, limitation of cytokine production is essential for the termination of inflammation and the prevention of endotoxic tissue damage (2).The production of inflammatory mediators is controlled at multiple levels, including transcription, translation, and protein stability (3). LPS promotes the nuclear import of NF-B (4) by indirectly regulating the activity of the IB kinase (IKK) complex, which phosphorylates IB␣, a factor that normally retains NF-B in the cytoplasm (5-7). Phosphorylated IB␣ is rapidly destroyed, thereby releasing NF-B to translocate to the nucleus and activate the transcription of target genes (8, 9). A proximal upstream LPSactivated factor is the Toll-like receptor, whose signaling pathway includes transforming growth factor beta-activated kinase 1 (TAK1), a mitogen-activated protein (MAP) kinase kinase kinase (10). TAK1 stimulation of p38 MAP kinase leads to IKK activity and NF-B import. LPS stimulation of TAK1 also triggers the stabilization of many mRNAs, which is dependent upon the interplay of several 3= untranslated region (UTR)-binding proteins such as those that associate with the AU-rich element (ARE) ...

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Section: Discussionsupporting

confidence: 85%

CPEB Regulation of TAK1 Synthesis Mediates Cytokine Production and the Inflammatory Immune Response

Ivshina

Alexandrov

Vertii

et al. 2015

Molecular and Cellular Biology

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“…Since primary cells respond earlier to LPS, their induction was reduced to 2 h. 24 Upon LPS stimulation wt-BMDM displayed a strong activation of the kinases p38 and ERK 1/2 after 10 min (Fig. 3A lane 2) that was clearly diminished in TLR4 -/--BMDM (Fig.…”

Section: Analysis Of the Lps Response In Primary Macrophagesmentioning

confidence: 94%

“…24 For deep sequencing, small RNA libraries were generated from isolated total RNA. MiRNAs differentially expressed in response to LPS were identified and classified through integration of the miR-Intess TM platform and miRBase libraries.…”

Section: Identification Of Differentially Expressed Mirnas In Lps-actmentioning

confidence: 99%

miR-155 targets Caspase-3 mRNA in activated macrophages

et al. 2015

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To secure the functionality of activated macrophages in the innate immune response, efficient life span control is required. Recognition of bacterial lipopolysaccharides (LPS) by toll-like receptor 4 (TLR4) induces downstream signaling pathways, which merge to induce the expression of cytokine genes and anti-apoptotic genes. MicroRNAs (miRNAs) have emerged as important inflammatory response modulators, but information about their functional impact on apoptosis is scarce. To identify miRNAs differentially expressed in response to LPS, cDNA libraries from untreated and LPSactivated murine macrophages were analyzed by deep sequencing and regulated miRNA expression was verified by Northern blotting and qPCR. Employing TargetScan TM we identified CASPASE-3 (CASP-3) mRNA that encodes a key player in apoptosis as potential target of LPSinduced miR-155. LPS-dependent primary macrophage activation revealed TLR4-mediated enhancement of miR-155 expression and CASP-3 mRNA reduction. Endogenous CASP-3 and cleaved CASP-3 protein declined in LPS-activated macrophages. Accumulation of miR-155 and CASP-3 mRNA in miRNA-induced silencing complexes (miRISC) was demonstrated by ARGONAUTE 2 (AGO2) immunoprecipitation. Importantly, specific antagomir transfection effectively reduced mature miR-155 and resulted in significantly elevated CASP-3 mRNA levels in activated macrophages. In vitro translation assays demonstrated that the target site in the CASP-3 mRNA 3'UTR mediates miR-155-dependent Luciferase reporter mRNA destabilization. Strikingly, Annexin V staining of macrophages transfected with antagomir-155 and stimulated with LPS prior to staurosporine (SSP) treatment implied that LPS-induced miR-155 prevents apoptosis through CASP-3 mRNA down-regulation. In conclusion, we report that miR-155-mediated CASP-3 mRNA destabilization in LPS-activated RAW 264.7 macrophages suppresses apoptosis, as a prerequisite to maintain their crucial function in inflammation.

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“…2C). Regulatory hnRNP-K-mRNA interactions have been demonstrated for the 5′ UTR (Evans et al, 2003) and 3′UTR (Liepelt et al, 2014;Naarmann et al, 2008;Ostareck et al, 1997). Protein class annotation using PANTHER allocated five proteins encoded by the 23 mRNAs to cytoskeletal proteins (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…NMHC IIA mRNA 5′UTR, 3′UTR and 3′UTR fragments were PCR amplified from K562 cDNA and cloned into pBSIIKS(+) (Stratagene, La Jolla, CA, USA) and pBSIIKS-Luc-pA-NB (Liepelt et al, 2014) using SpeI (5′UTR) and HindIII (3′UTR), to generate pBSIIKS-NMHC IIA-5′UTR, pBSIIKS-NMHC IIA-3′UTR, pBSIIKS-NMHC IIA-3′F1, pBSIIKS-NMHC IIA-3′F2, pBSIIKS-NMHC IIA-3′F3, pBSIIKS-Luc-NMHC IIA-3′F1-pA and pBSIIKS-Luc-NMHC IIA-3′F3-pA. For pBSIIKS-Luc-NMHC IIA-3′F1 S1_M2)-pA and pBSIIKS-Luc-NMHC IIA-3′F1(S2_M3)-pA, gene fragments (Eurofins Genomics, Table S10) corresponding to NMHC IIA-3′F1 with nucleotide exchanges were amplified and cloned into pBSIIKS-Luc-pA-NB. pBSIIKS-Luc-NMHC IIA-3′F1_ΔS1-pA, pBSIIKSLuc-NMHC IIA-3′F1_ΔS2-pA and pBSIIKS-Luc-NMHC IIA-3′F1_ΔS1, S2-pA were generated by site-directed mutagenesis from pBSIIKS-Luc-NMHC IIA-3′F1-pA.…”

Section: Plasmidsmentioning

confidence: 99%

Translational control mediated by hnRNP K links NMHC IIA to erythroid enucleation

Vries

Brendle

Bähr-Ivacevic

et al. 2016

Journal of Cell Science

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Post-transcriptional regulation is crucial for structural and functional alterations in erythropoiesis. Enucleation of erythroid progenitors precedes reticulocyte release into circulation. In enucleated cells, reticulocyte 15-lipoxygenase (r15-LOX, also known as ALOX15) initiates mitochondria degradation. Regulation of r15-LOX mRNA translation by hnRNP K determines timely r15-LOX synthesis in terminal maturation. K562 cells induced for erythroid maturation recapitulate enucleation and mitochondria degradation. HnRNP K depletion from maturing K562 cells results in enhanced enucleation, which even occurs independently of maturation. We performed RIP-Chip analysis to identify hnRNP K-interacting RNAs comprehensively. Non-muscle myosin heavy chain (NMHC) IIA (also known as MYH9) mRNA co-purified with hnRNP K from noninduced K562 cells, but not from mature cells. NMHC IIA protein increase in erythroid maturation at constant NMHC IIA mRNA levels indicates post-transcriptional regulation. We demonstrate that binding of hnRNP K KH domain 3 to a specific sequence element in the NMHC IIA mRNA 3′UTR mediates translation regulation in vitro. Importantly, elevated NMHC IIA expression results in erythroidmaturation-independent enucleation as shown for hnRNP K depletion. Our data provide evidence that hnRNP-K-mediated regulation of NMHC IIA mRNA translation contributes to the control of enucleation in erythropoiesis.

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Translation control of TAK1 mRNA by hnRNP K modulates LPS-induced macrophage activation

Cited by 30 publications

References 61 publications

CPEB Regulation of TAK1 Synthesis Mediates Cytokine Production and the Inflammatory Immune Response

CPEB Regulation of TAK1 Synthesis Mediates Cytokine Production and the Inflammatory Immune Response

miR-155 targets Caspase-3 mRNA in activated macrophages

Translational control mediated by hnRNP K links NMHC IIA to erythroid enucleation

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