Ultrastructural Localization of Interferon-Inducible Double-Stranded RNA-Activated Enzymes in Human Cells

Besse, Sylvie; Rebouillat, Dominique; Marié, Isabelle; Puvion‐Dutilleul, Francine; Ag, Hovanessian

doi:10.1006/excr.1997.3908

Cited by 57 publications

(45 citation statements)

References 76 publications

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“…However, electron micrograph analysis and immunogold labeling has also indicated that a fraction of PKR resides in the nucleus (48,49). Since the NFARs have a consensus bipartite nuclear localization signal (Fig.…”

Section: Isolation and Identification Of Nfar-1 And -2-mentioning

confidence: 99%

Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, That Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR

Saunders

Perkins

Balachandran

et al. 2001

Journal of Biological Chemistry

114

148

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We report here the isolation and characterization of two proteins, NFAR-1 and -2, which were isolated through their ability to interact with the dsRNA-dependent protein kinase, PKR. The NFAR proteins, of 90 and 110 kDa, are derived from a single gene through alternative splicing and are evolutionarily conserved nuclear phosphoproteins that interact with doublestranded RNA. Both NFAR-1 and -2 are phosphorylated by PKR, reciprocally co-immunoprecipitate with PKR, and colocalize with the kinase in a diffuse nuclear pattern within the cell. Transfection studies indicate that the NFARs regulate gene expression at the level of transcription, probably during the processing of pre-mRNAs, an activity that was increased in fibroblasts lacking PKR. Subsequent functional analyses indicated that amino acids important for NFAR's activity were localized to the C terminus of the protein, a region that was found to specifically interact with FUS and SMN, proteins also known as regulators of RNA processing. Accordingly, both NFARs were found to associate with both pre-mRNAs and spliced mRNAs in post-transcriptional studies, similar to the known splicing factor ASF/ SF-2. Collectively, our data indicate that the NFARs may facilitate double-stranded RNA-regulated gene expression at the level of post-transcription and possibly contribute to host defense-related mechanisms in the cell.

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Section: Isolation and Identification Of Nfar-1 And -2-mentioning

confidence: 99%

Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, That Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR

Saunders

Perkins

Balachandran

et al. 2001

Journal of Biological Chemistry

114

148

View full text Add to dashboard Cite

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“…ribosomes in the cytoplasm and the rough endoplasmic reticulum (MacQuillan et al, 2009) but also localises to the juxta-nuclear cytoplasmic layer, the nuclear envelope and within the nucleus itself (Jeffrey et al, 1995;Besse et al, 1998). Localisation of active PKR within the cytoplasm is necessary for access to its substrates such as eIF2 associated with the ribosomes (Zhu et al, 1997) and the IKK complex that releases IB, the inhibitor of kappa Bto allow the translocation of nuclear factor kappa B (NFB) to the nucleus (Kumar et al, 1994).…”

Section: Sj Gilbert Et Al Pkr Mediates Osm and Il-1 Signalling In Chmentioning

confidence: 99%

“…Interferon treatment of peripheral blood mononuclear cells for 24 h increases the amount of active PKR in the nucleus (MacQuillan et al, 2009). The role of PKR in the nucleus is less well defi ned but may involve ribosome biogenesis (Jimenez-Garcia et al, 1993), regulation of the amount of RNA molecules released into the cytoplasm (Besse et al, 1998), and enhancement of TNF precursor transcript splicing (Osman et al, 1999). Protein from an equivalent cell number was analysed by Western blotting for type II collagen (n = 4 replicates per treatment, representative data shown from 2 independent experiments).…”

Section: Sj Gilbert Et Al Pkr Mediates Osm and Il-1 Signalling In Chmentioning

confidence: 99%

Protein kinase R plays a pivotal role in oncostatin M and interleukin-1 signalling in bovine articular cartilage chondrocytes

Gilbert

Blain²,

Al‐Sabah³

et al. 2012

eCM

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This study investigated whether treatment of articular cartilage chondrocytes with a combination of oncostatin M (OSM) and interleukin-1 (IL-1) could induce a degradative phenotype that was mediated through the protein kinase R (PKR) signalling pathway. High-density monolayer cultures of full depth, bovine chondrocytes were treated with a combination of OSM and IL-1 (OSM+IL-1) for 7 days. To inhibit the activation of PKR, a pharmacological inhibitor of PKR was added to duplicate cultures. Pro-and active matrix metalloproteinase-9 (MMP9) and MMP9 mRNA were signifi cantly upregulated by OSM+IL-1 through a PKR dependent mechanism. ADAMTS4 and ADAMTS5 mRNA were also upregulated by OSM+IL-1. The upregulation of ADAMTS4 and ADAMTS5 were, in part, mediated through PKR. OSM+IL-1 resulted in a loss of type II collagen, which could not be rescued by PKR inhibition. OSM+IL-1 reduced the expression of COL2A1 (type II collagen), COL9A1 (type IX collagen), COL11A1 (type XI collagen), and ACAN (aggrecan) mRNAs. Expression of type II and XI collagen and aggrecan was reduced further when PKR was inhibited. OSM+IL-1 resulted in an 11-fold increase in TNFα mRNA which was, in part, mediated through the PKR pathway. This study demonstrates, for the fi rst time, that a number of catabolic and pro-infl ammatory effects known to be important in human arthritis and induced by OSM and IL-1, are mediated by the PKR signalling pathway.Keywords: Articular cartilage, chondrocytes, protein kinase R, oncostatin M, interleukin-1, matrix metalloproteinase, aggrecanase, tumour necrosis factor alpha, type II collagen.

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“…Yanase and coworkers have shown that during IFNα induced apoptosis there is a release of cytochrome c from mitochondria, a decline in mitochondrial membrane potential and caspase 3 activation [121]. The different partners of the 2-5A pathway are localized in mitochondria: RNase L, RLI and p69 form of 2-5A-synthetase [100,122,123]. This pathway is induced by IFNα treatment and RNase L participates in mitochondrial mRNA degradation.…”

Section: ) Apoptosismentioning

confidence: 99%

Diverse functions of RNase L and implications in pathology

Bisbal

Silverman

2007

Biochimie

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The endoribonuclease L (RNase L) is the effector of the 2-5A system, a major enzymatic pathway involved in the molecular mechanism of interferons (IFN). RNase L is a very unusual nuclease with a complex mechanism of regulation. It is a latent enzyme, expressed in nearly every mammalian cell type. Its activation requires its binding to a small oligonucleotide, 2-5A. 2-5A is a series of unique 5′-triphosphorylated oligoadenylates with 2′-5′ phosphodiester bonds. By regulating viral and cellular RNA expression, RNase L plays an important role in the antiviral and antiproliferative activities of IFN and contributes to innate immunity and cell metabolism. The 2-5A/RNase L pathway is implicated in mediating apoptosis in response to viral infections and to several types of external stimuli. Several recent studies have suggested that RNase L could have a role in cancer biology and evidence of a tumor suppressor function of RNase L has emerged from studies on the genetics of hereditary prostate cancer. KeywordsInterferon; RNase L; RNA expression; apoptosis; prostate; virus; cancer I) IntroductionThe endoribonuclease L (RNase L) is the effector of the 2-5A system, a major enzymatic pathway regulated by interferons (IFN) [1] (Figure 1). The 2-5A system is one of the two antiviral pathways induced by IFN and activated by double stranded RNA (dsRNA), the other is mediated by the dsRNA dependent protein kinase (PKR) [2]. RNase L is a very unusual nuclease. It is a latent enzyme, expressed in nearly every mammalian cell type. Its activation requires its binding to a small oligonucleotide, 2-5A ( Figure 1). 2-5A itself is very unusual, consisting of a series of 5′-triphosphorylated oligoadenylates with 2′-5′ phosphodiester bonds in contrast to the 3′-5′ linkages found in RNA and DNA. The initial and essential observation was made by Ian Kerr's group in 1974 reporting an IFN-induced increase in the sensitivity of protein synthesis to inhibition by dsRNA [3]. Peter Lengyel's group observed increased nuclease activity in extracts of interferon treated cells incubated with dsRNA [4,5]. The identification by Ian Kerr's group of the activators of this nuclease, 2-5A [6] and of the enzyme responsible for their synthesis, the 2-5A-synthetase [7][8][9], led to the discovery of the 2-5A pathway. Clemens and Williams directly demonstrated a nuclease, now recognized as RNase Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. (Figure 2). The N-terminal domain could be considered as the regulatory domain of RNase L. It is composed of eight complete and one partial ankyrin motifs (R1-R9). Two wal...

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Ultrastructural Localization of Interferon-Inducible Double-Stranded RNA-Activated Enzymes in Human Cells

Cited by 57 publications

References 76 publications

Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, That Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR

Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, That Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR

Protein kinase R plays a pivotal role in oncostatin M and interleukin-1 signalling in bovine articular cartilage chondrocytes

Diverse functions of RNase L and implications in pathology

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