Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An.

Slattery, Elizabeth; Ghosh, Nimai K.; Samanta, Himadri; Lengyel, Péter

doi:10.1073/pnas.76.10.4778

Cited by 132 publications

(40 citation statements)

References 22 publications

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“…The radiobinding assay is based on the high affinity of ppp(A2'p),A for a ppp(A 2'p),A-binding protein which is thought to be the ppp(A2'p)"A-dependent RNase [9,12,25,281. Here the assay has been used to compare the affinities of different analogues by determining the concentration of analogue required to displace ppp(A2'p),A-[32P]pCp from the binding protein (presumptive RNase) which is retained on nitrocellulose filters.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, unlike others [29, 301, we have been unable, to date, to obtain any effect of 'core' or its analogues on intact cells which have not been subjected to prior permeabilisation [ 3 3 ] . ('Cores' are not efficiently precipitated using the calcium phosphate tech- (3,5,7,9) 200 nM, tracks (4,6,8,10) 1 pM of (3,4) p(A2'p),A, (5, 6) p(A2'p),A2',3'CH2, (7,8) p(A2'p), A3'0CH3 and (9,10) p(A2'p),dA. Incubations were for 2 h at 30':C (Materials and Methods).…”

Section: Discussionmentioning

confidence: 99%

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Analogues and Analogue Inhibitors of ppp(A2′p)_nA

Haugh¹,

Cayley²,

Serafinowska³

et al. 1983

European Journal of Biochemistry

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Higher Oligomers of ppp(A2′p)n together with (A2′p)nA, (A2′p)2A3′OCH3, (A2′p)2A2′,3′CH2, (A2′p)2dA, (dA2′p)2dA, their 5′‐monophosphates and 5′‐S‐methylphorothioates have been investigated for relative stability and biological activity in mouse and human cells and mouse, human, and rabbit cell‐free systems. The oligomers from trimer to heptamer inhibited protein and DNA synthesis when introduced into intace mouse cells and activated the ppp(A2′p)n A‐dependent RNase at below nanomolar concentrations in mouse cell extracts. The 5′‐diphosphates pp(A2′p)2A and corresponding analogues were active both in cell‐free systems and on introduction into intact cells. The exception to this was the all 3′‐deoxyadenosine analogue pp(dA2′p)2 dA which failed to activate the ppp(A2′p)nA‐dependent nuclease in the mouse L and human (Daudi and HeLa) cell extracts tested. Of the active analogues the 3′‐OCH3 appeared to be the most stable in the cells and systems employed. On the other hand the non‐phosphorylated ‘core’ (A2′p)2A and its 3′‐sunstituted analogues were inactive in mouse L and Ehrlich acites tumours cell‐free systems and had to effect on intact (non‐permeabilised) 3T3 cells. The 5′‐monophoshate, p(A2′p)2A, can act as an inhibitor of ppp(A2′p)nA in mouse cell‐free systems [Torrence, Imai and Johnston (1981) Proc. Natl Acad. Sci. USA, 78, 5993‐59997]. Here in intact mouse L cells or extracts from interferon‐treated human (Daudi) cells, however, it mimicked the action of ppp(A2′p)2A, possibly through conversion to the 5′‐diphosphate or 5′‐S‐methylphosphorothioate derivatives of the 3′‐substituted analogues are both more stable to exonucleolytic cleavage and unlikely to be converted to the 5′‐diphosphated or 5′‐triphosphates. They are analogue inhibitors of ppp(A2′p)nA in mouse L cell extracts. How widely they will be effective in a variety of cell‐free systems and intact cells remains to be established. The 5′‐diphosphate pp(A2′p)2A and corresponding analogues were not equally active, nor was the CH3Sp(A2′p)2A2′,3′CH2 equally effective as an analogue inhibitor, in different cell‐free systems. This emphasises the apparent differences in the properdties of the ppp(A2′p)nA‐dependent RNases from different sources. Accordingly, in looking for a generally effective analogue inhibitor of ppp(A2′p)2A its activity in a variety of extracts should be tested and in any search for further analogues for potential clincial use human cells and extracts should be employed.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analogues and Analogue Inhibitors of ppp(A2′p)_nA

Haugh¹,

Cayley²,

Serafinowska³

et al. 1983

European Journal of Biochemistry

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“…RNase L was first described as a 185 kDa complex by gel filtration experiments [18] then as a 78-80 kDa protein in gel denaturing conditions [19,20]. These two forms seem to coexist in cells, their ratio depending on the cell type and experimental procedures use to detect RNase L [21][22][23].…”

Section: Ii) Rnase L Structurementioning

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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“…(a) The kinetics of the induction of (2'-5')A, synthetase [9] and in some cases the kinetics of the induction of the dsRNA-dependent protein kinase activity [I21 correlate with the development of the antiviral effect. (b) The activation of endoribonuclease F (RNase F) by (2'-5')A, [6,13,14] and the inactivation of eIF-2 by phosphorylation of its x subunit are believed to be involved with the inhibition of protein synthesis in interferontreated cells [I 5,161; moreover, the addition of exogenous eIF-2 [17] as well as the dephosphorylation of PI and of the x subunit of eIF-2 has been associated with the reversal of dsRNA, double-stranded R N A ; (2'-5')A,, a mixturc of 5'-triphosphoadenyl Z'-S'-linked oligoadenylates; (2'-5')A3, 5'-triphosphoadenyl-(2'-5')adenylyl(2'-5')adenosine; PI, the interferon-induced 67000-dalton protcin; eIF-2, eukaryotic initiation factor 2; RNase F, the (2'-5')A,,-dependent ribonuclease; EMCV, encephaloinyocarditis virus; VSV, vesicular stomatitis virus; MMuLV, Moloney murine leukemia virus; Hepes, 4-(2-hydroxyl)-l-piperazineethanesulfonic acid; BMK, baby hamster kidney; Mes, 4-morpholineethanesutfonic acid.…”

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confidence: 99%

A Mouse Cell Line, which Is Unprotected by Interferon against Lytic Virus Infection, Lacks Ribonuclease F Activity

Epstein

Czarniecki

Jacobsen

et al. 1981

European Journal of Biochemistry

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A mouse cell line, N I H 3T3, does not respond to some of the activities of interferon. Even after treatment with high concentrations of interferon the replication of lytic viruses, such as encephalomyocarditis virus (EMCV) and vesicular stomatitis virus (VSV) is not inhibited in these cells. In contrast, interferon treatment of these same cells results in the inhibition of Moloney murine leukemia virus (MMuLV) production. We have analyzed enzymatic pathways which are induced by interferon in these cells. After interferon treatment, the level of the (2'-5')oligoadenylate [(2'-5')A,] synthetase activity and the phosphorylation of the 67000-dalton protein (PI) are enhanced in NIH 3T3 cells to approximately the same level as interferon-sensitive mouse L-cells. Moreover, NIH 3T3 and L-cells contain approximately the same levels of enzymcs which inactivate (2'-5')A,. Both exogenously added (2'-5')Aj or double-stranded RNA (ds RNA) failed to inhibit protein synthesis in NIH 3T3 extracts even though they were potent inhibitors of L-cell extract-directed protein synthesis. Direct measurements of the (2'-S')A,-dependent ribonuclease F (RNase F) failed to detect such activity in NIH 3T3 cells. Our results, therefore, suggest that the presence of RNase F activity is necessary for the interferon-induced antiviral activity against EMCV and against VSV. The induction of protein kinase activity by interferon treatment of NIH 3T3 cells appears to have no direct effect on EMCV and VSV replication.

show abstract

Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An.

Abstract: Among the mediators of interferon action are one enzyme that is activated by double-stranded RNA to convert

Cited by 132 publications

References 22 publications

Analogues and Analogue Inhibitors of ppp(A2′p)_nA

Analogues and Analogue Inhibitors of ppp(A2′p)_nA

Diverse functions of RNase L and implications in pathology

A Mouse Cell Line, which Is Unprotected by Interferon against Lytic Virus Infection, Lacks Ribonuclease F Activity

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Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An.

Abstract: Among the mediators of interferon action are one enzyme that is activated by double-stranded RNA to convert

Cited by 132 publications

References 22 publications

Analogues and Analogue Inhibitors of ppp(A2′p)nA

Analogues and Analogue Inhibitors of ppp(A2′p)nA

Diverse functions of RNase L and implications in pathology

A Mouse Cell Line, which Is Unprotected by Interferon against Lytic Virus Infection, Lacks Ribonuclease F Activity

Contact Info

Product

Resources

About

Analogues and Analogue Inhibitors of ppp(A2′p)_nA

Analogues and Analogue Inhibitors of ppp(A2′p)_nA