Biochemical Evidence for a Novel Low Molecular Weight 2-5A-Dependent RNase L in Chronic Fatigue Syndrome

Suhadolnik, Robert J.; Peterson, Daniel L.; O’Brien, Kerry L.; Cheney, Paul R.; Herst, C.v.; Reichenbach, Nancy L.; Kon, Ning; Horvath, Susan E.; Iacono, Kathryn T.; Adelson, Martin E.; Meirleir, Kenny De; Becker, Pascale De; Charubala, Ramamurthy; Pfleiderer, Wolfgang

doi:10.1089/jir.1997.17.377

Cited by 99 publications

(43 citation statements)

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“…Several reports indicated the up-regulation of components of the 2-5A/RNase L pathway in extracts of peripheral blood mononuclear cells (PBMCs) from CFS patients [145,146] as well as the accumulation of a low molecular weight 2-5A-binding protein of 37 kDa [147]. It has been proposed that this polypeptide could be a biochemical marker for CFS [148,149].…”

Section: ) Chronique Fatigue Syndrome (Cfs)mentioning

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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Section: ) Chronique Fatigue Syndrome (Cfs)mentioning

confidence: 99%

Diverse functions of RNase L and implications in pathology

Bisbal

Silverman

2007

Biochimie

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“…Khalid Khabar showed that RNase L could affect the stability of PKR mRNA and levels of PKR, demonstrating that the two pathways are in a type of balance [56]. Studies by the groups of Robert Suhadolnik [57,58] and Bernard LeBleu [59,60] described up-regulation of the 2-5A system and accumulation of a 37 kDa polypeptide from RNase L in PBMC of patients with chronic fatigue syndrome. In addition, Lawrence Kuo's group at Merck Research Laboratories performed outstanding enzymology and biophysical studies of RNase L [61][62][63][64].…”

Section: Molecular Cloning Of Rnase Lmentioning

confidence: 99%

A scientific journey through the 2-5A/RNase L system

Silverman

2007

Cytokine & Growth Factor Reviews

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The antiviral and antitumor actions of interferons are caused, in part, by a remarkable regulated RNA cleavage pathway known as the 2-5A/RNase L system. 2′-5′ linked oligoadenylates (2-5A) are produced from ATP by interferon-inducible synthetases. 2-5A activates pre-existing RNase L, resulting in the cleavage of RNAs within single-stranded regions. Activation of RNase L by 2-5A leads to an antiviral response, although precisely how this happens is a subject of ongoing investigations. Recently, RNase L was identified as the hereditary prostate cancer 1 gene. That finding has led to the discovery of a novel human retrovirus, XMRV. My scientific journey through the 2-5A system recounts some of the highlights of these efforts. Knowledge gained from studies on the 2-5A system could have an impact on development of therapies for important viral pathogens and cancer.Keywords 2-5A; RNase L; interferon; cancer; virus Background on the 2-5A/RNase L systemOver the past thirty-years, investigations into the mechanisms of interferon (IFN) action have elucidated how antiviral innate immunity operates within and between mammalian cells. The focus of my work over this period has been, and continues to be, probing the biology and biochemistry of the 2-5A/RNase L system, and studying its roles in health and disease. My scientific journey, from basic research to clinical studies, are summarized in this monograph. The 2-5A/RNase L system is one the principal pathways by which IFNs suppress viral infections. Exposure of cells to IFNs induces expression of genes that result in an "antiviral state". Type I IFNs bind to the IFN-α receptor 1 (IFNAR1) and IFNAR2 polypeptide chains on cell surfaces initiating JAK-STAT signaling to the IFN stimulated genes (ISGs) [1]. Included among over a hundred different ISGs are genes encoding 2-5A synthetases (OAS) [2,3]. The OAS genes are a family of ISGs that function in the 2-5A/RNase L system (Fig. 1). In humans there are three functional OAS genes (OAS1-3), resulting in 8 to 10 OAS isoforms due to alternative mRNA splicing [4]. In mice, in addition to OAS2 and OAS3 there are 7 separate OAS1 genes, including OAS1b, the flavivirus resistance gene (Flv r ] [5][6][7][8]. When stimulated by dsRNA, the functional OAS proteins produce a series of short 5′-phosphorylated, 2′,5′-linked oligoadenylates collectively referred to as 2-5A [p x 5′A(2′p5′A) n ; x = 1-3; n≥2] from ATP [9]. The first molecular clone for an OAS was obtained by Michel Revel's lab [10]. Biochemical characterization of OAS proteins by Ara Hovanessian's lab [11][12][13][14] and Ganes Sen's lab [15][16][17][18][19] and a crystal structure by Rune Hartmann and Vivien Yee (in collaboration with Ganes Sen and Just Justesen) [20] have led to functional and structural insight into the OAS family of 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, type...

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“…Previous studies from this laboratory (1) described structural similarities between two forms of 2Ј,5Ј-oligoadenylate (2-5A) 1 dependent RNase L in extracts of peripheral blood mononuclear cells (PBMC). The 2-5A-dependent 80-kDa RNase L is the terminal enzyme in the 2-5A synthetase/RNase L antiviral defense pathway (2).…”

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Characterization of a 2′,5′-Oligoadenylate (2–5A)-dependent 37-kDa RNase L

Shetzline¹,

Suhadolnik

2001

Journal of Biological Chemistry

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Upregulation of key components of the 2,5-oligoadenylate (2-5A) synthetase/RNase L pathway have been identified in extracts of peripheral blood mononuclear cells from individuals with chronic syndrome, including the presence of a low molecular weight form of RNase L. In this study, analysis of 2,5-Oligoadenylate (2-5A) binding and activation of the 80-and 37-kDa forms of RNase L has been completed utilizing photolabeling/immunoprecipitation and affinity assays, respectively. Saturation of photolabeling of the 80-and the 37-kDa RNase L with the 2-5A azido photoprobe, [ Previous studies from this laboratory (1) described structural similarities between two forms of 2Ј,5Ј-oligoadenylate (2-5A) 1 -dependent RNase L in extracts of peripheral blood mononuclear cells (PBMC). The 2-5A-dependent 80-kDa RNase L is the terminal enzyme in the 2-5A synthetase/RNase L antiviral defense pathway (2). RNase L is distinguished from other RNases in that it requires 2-5A for allosteric activation (reviewed in Refs. 1 and 3-6). The 80-kDa RNase L plays an essential role in the inhibition of viral mRNA and thus the inhibition of viral protein synthesis (3, 4). In addition to the 80-kDa form of RNase L, a 40/42-kDa form has been identified and is thought to be a proteolytic degradation product of the 80-kDa RNase L (7, 8).As part of ongoing studies on the up-regulation of the 2-5OAS/RNase L antiviral pathway, a previously unknown 37-kDa form of RNase L has recently been identified in extracts of PBMC from individuals who met the diagnostic criteria for chronic fatigue syndrome (CFS) (9 -12). We had previously demonstrated that the 2-5OAS/RNase L pathway is constitutively active in PBMC extracts from individuals with CFS; 2-5OAS is predominantly in its activated form and levels of bioactive 2-5A and RNase L enzyme activity are elevated in CFS PBMC extracts compared with healthy control PBMC extracts (13,14). Continued analyses of CFS PBMC extracts have revealed the presence of a novel low molecular mass RNase L that binds 2-5A, exhibits 2-5A-dependent RNase L activity, and has a molecular mass of 37-kDa under denaturing conditions (1, 10). The 37-kDa RNase L has not been observed in PBMC extracts from individuals diagnosed with fibromyalgia or depression (15).Two subsets of individuals with CFS have been identified in PBMC extracts using azido photoaffinity labeling with [ 32 P]pApAp(8-azidoA) and immunoprecipitation with a polyclonal antibody specific to the human 80-kDa RNase L (1, 10). One subset contains only the 37-kDa RNase L and the second subset contains both the 80-and 37-kDa RNase L. The subset of CFS PBMC containing only the 37-kDa RNase L had the higher level of RNase L activity compared with the subset containing both the 80-and 37-kDa forms of RNase L (2). In a two-site randomized-coded study, the presence of the 37-kDa RNase L also correlated with the level of clinical disability experienced by the study subjects (10).The elucidation of the molecular structure of the 37-kDa RNase L has been a major effort in this laborat...

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Biochemical Evidence for a Novel Low Molecular Weight 2-5A-Dependent RNase L in Chronic Fatigue Syndrome

Cited by 99 publications

References 36 publications

Diverse functions of RNase L and implications in pathology

Diverse functions of RNase L and implications in pathology

A scientific journey through the 2-5A/RNase L system

Characterization of a 2′,5′-Oligoadenylate (2–5A)-dependent 37-kDa RNase L

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