Regulation of<i>in vitro</i>translation by double-stranded RNA in mammalian cell mRNA preparations

Pratt, Gerg; Galpine, Angela R.; Sharp, Nigel A.; Palmer, Susan; Clemens, Michael J.

doi:10.1093/nar/16.8.3497

Cited by 20 publications

(9 citation statements)

References 44 publications

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“…The latter inhibits PKR but has no effect on the activation of 2-5A synthetase by dsRNA. Thus, 2-aminopurine can be used to distinguish between inhibition of translation caused by PKR and that caused by the 2-5A/RNase L system (31,32). The abilities of various concentrations of the sx112 aptamer to inhibit protein synthe- FIG.…”

Section: Resultsmentioning

confidence: 99%

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Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Hartmann¹,

Nørby²,

Martensen³

et al. 1998

Journal of Biological Chemistry

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A number of small RNA molecules that are high affinity ligands for the 46-kDa form of human 2-5 oligoadenylate synthetase have been identified by the SELEX method. Surface plasmon resonance analysis indicates that these RNAs bind to the enzyme with dissociation constants in the nanomolar range. Competition experiments indicate that the binding site for the small RNAs on the 2-5 oligoadenylate synthetase molecule at least partially overlaps that for the synthetic doublestranded RNA, poly(I)⅐poly(C). Several of the RNAs function as potent activators of 2-5 oligoadenylate synthetase in vitro, although there is no correlation between binding affinity and ability to activate. The RNA aptamers having the strongest activation potential appear to have few base-paired regions. This suggests that 2-5 oligoadenylate synthetase, which has previously been believed to be activated only by double-stranded RNA, can also be activated by RNA ligands with little secondary structure. Since 2-5 oligoadenylate synthetase possesses no homology to other known RNA-binding proteins, the development of small specific ligands by SELEX should facilitate studies of RNA-protein interactions and may reveal novel features of the structurefunction relationships involving this enzyme. Double-stranded RNA (dsRNA) 1 binds to and activates a number of interferon-induced proteins, namely the family of 2Ј-5Ј oligoadenylate synthetases (2-5A synthetase) and the protein kinase PKR (reviewed in Refs.

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Section: Resultsmentioning

confidence: 99%

“…RNA-protein binding was assayed by the nitrocellulose filter method. In all experiments, 32 P-labeled sx112 RNA was present at a final concentration of 6 nM (0.2 g/ml). All binding reactions contained 0.2 mg/ml BSA.…”

Section: Figmentioning

confidence: 99%

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Hartmann¹,

Nørby²,

Martensen³

et al. 1998

Journal of Biological Chemistry

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“…Inhibition of protein synthesis may take place in a localized fashion within the cell (17). It is noteworthy to mention that another research group has previously detected the existence of dsRNA structures in RNA preparations of certain cell lines by its ability to activate in vitro phosphorylation of eIF-2ac (59). The biological relevance of these structures is underlined by the fact that genes with potential cytokine activity and identified as platelet-derived growth factor inducible were also shown to be regulated by dsRNA (63,75 different pathway than when platelet-derived growth factor is the inducer (26).…”

Section: Resultsmentioning

confidence: 99%

Action of spontaneously produced beta interferon in differentiation of embryonal carcinoma cells through an autoinduction mechanism.

Belhumeur¹,

Lanoix²,

Blais³

et al. 1993

Mol. Cell. Biol.

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“…Previously, such long, intramolecular, rod-like structures have been observed in plant viroids and viruses such as hepatitis delta virus, but the molecules we observed are within cellular RNA. In mammals, viral dsRNA activates dsRNA binding proteins involved in the interferon response, and it has been speculated that cells have dsRNA that can activate these proteins in the absence of viral infection (24)(25)(26)(27)(28). Possibly, C. elegans also has mechanisms for regulation by dsRNA, as suggested by the recent discovery of RNA interference (29).…”

Section: Discussionmentioning

confidence: 99%

Long RNA hairpins that contain inosine are present in Caenorhabditis elegans poly(A) ⁺ RNA

Morse

Bass

1999

Proc. Natl. Acad. Sci. U.S.A.

137

113

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Adenosine deaminases that act on RNA (ADARs) are RNA-editing enzymes that convert adenosine to inosine within double-stranded RNA. In the 12 years since the discovery of ADARs only a few natural substrates have been identified. These substrates were found by chance, when genomically encoded adenosines were identified as guanosines in cDNAs. To advance our understanding of the biological roles of ADARs, we developed a method for systematically identifying ADAR substrates. In our first application of the method, we identified five additional substrates in Caenorhabditis elegans. Four of those substrates are mRNAs edited in untranslated regions, and one is a noncoding RNA edited throughout its length. The edited regions are predicted to form long hairpin structures, and one of the RNAs encodes POP-1, a protein involved in cell fate decisions.Adenosine deaminases that act on RNA (ADARs) have been detected in every metazoan examined (reviewed in refs. 1 and 2), and cDNAs encoding various members of the enzyme family have been cloned (3). The product of adenosine deamination, inosine, is found within mRNA in tissue-specific amounts that correlate with the amounts of ADAR in various tissues (4), and the observed levels suggest many mRNAs are acted on by ADARs. Yet, in the 12 years since the discovery of ADARs (5-8), only a few natural substrates have been identified (reviewed in ref.2). These substrates have been found entirely by chance, when genomically encoded adenosines were identified as guanosines in cDNAs. Such A to G transitions are diagnostic of A to I conversions, because inosine prefers to pair with cytidine and is changed to a G during cDNA synthesis.Studies of the serendipitously discovered substrates show that one function of ADARs is to deaminate adenosines within codons. In this way, multiple protein isoforms can be synthesized from a single encoded mRNA. ADARs are involved in producing functionally important isoforms of mammalian serotonin receptors (9), several mammalian glutamate receptor subunits (10-12), and the virally encoded hepatitis delta antigen (13). In addition, A to G transitions have been detected within codons of several other viral and cellular transcripts where function has not yet been verified (reviewed in ref. 2).Many additional functions of ADARs have been proposed. Conceivably, A to I conversions could affect any process that involves sequence-specific interactions, so effects on RNA processing, stability, and translatability are all possible. Further, because adenosine deamination can alter RNA structure, sequence-independent processes also could be affected. In fact, the intrinsic properties of ADARs suggest their primary and primordial functions remain to be elucidated. For example, ADARs act promiscuously on completely base-paired double-stranded RNA (dsRNA) substrates, deaminating Ϸ50% of the adenosines in a single molecule. Obviously, such rampant deamination is ill-suited for generating protein isoforms of precise function. In mRNAs where ADARs are known to produce ...

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Regulation ofin vitrotranslation by double-stranded RNA in mammalian cell mRNA preparations

Cited by 20 publications

References 44 publications

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Action of spontaneously produced beta interferon in differentiation of embryonal carcinoma cells through an autoinduction mechanism.

Long RNA hairpins that contain inosine are present in Caenorhabditis elegans poly(A) ⁺ RNA

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Regulation ofin vitrotranslation by double-stranded RNA in mammalian cell mRNA preparations

Cited by 20 publications

References 44 publications

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Activation of 2′-5′ Oligoadenylate Synthetase by Single-stranded and Double-stranded RNA Aptamers

Action of spontaneously produced beta interferon in differentiation of embryonal carcinoma cells through an autoinduction mechanism.

Long RNA hairpins that contain inosine are present in Caenorhabditis elegans poly(A) + RNA

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Long RNA hairpins that contain inosine are present in Caenorhabditis elegans poly(A) ⁺ RNA