Expression cloning of 2-5A-dependent RNAase: A uniquely regulated mediator of interferon action

2-5A-dependent RNAase, an interferon-induced enzyme that is activated by 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A), is implicated in both the molecular mechanisms of interferon action and the fundamental control of RNA stability in mammalian cells. Here we report the expression cloning and analysis of murine and human 2-5A-dependent RNAases. The 2-5A binding properties and RNAse activities of recombinant and naturally occurring forms of 2-5A-dependent RNAase were identical. Interferon induction of 2-5A-dependent RNAse expression was demonstrated by measuring the mRNA levels in cells treated with interferon and cycloheximide. Analysis of aligned murine and human 2-5A-dependent RNAse sequences revealed several intriguing features, including similarity to RNAase E, which is implicated in the control of mRNA stability in E. coli. Interestingly, a duplicated phosphate-binding loop motif was determined by deletion analysis and site-directed mutagenesis to function in the binding of 2-5A.

[1]  I. Kerr,et al.  Affinity labelling and characterization of the ppp(A2'p)nA-dependent endoribonuclease from different mammalian sources. , 2005, European journal of biochemistry.

[2]  G. Floyd-Smith.  (2′‐5′)An‐dependent endoribonuclease: Enzyme levels are regulated by IFNβ, IFNγ, and cell culture conditions , 1988 .

[3]  W. Kabsch,et al.  Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation , 1989, Nature.

[4]  M. Vigneron,et al.  Constitutive expression of (2′–5′) oligo A synthetase confers resistance to picornavirus infection , 1987, Nature.

[5]  A. Miczak,et al.  The gene specifying RNase E (rne) and a gene affecting mRNA stability (ams) are the same gene , 1991, Molecular microbiology.

[6]  R. Silverman,et al.  Purification and analysis of murine 2-5A-dependent RNase. , 1988, The Journal of biological chemistry.

[7]  T. Hunter,et al.  The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. , 1988, Science.

[8]  A. Hovanessian,et al.  The 69-kDa 2-5A synthetase is composed of two homologous and adjacent functional domains. , 1992, The Journal of biological chemistry.

[9]  G. Stark,et al.  2-5A synthetase: assay, distribution and variation with growth or hormone status , 1979, Nature.

[10]  M. I. Johnston,et al.  Purification and characterization of a 2'-phosphodiesterase from bovine spleen. , 1987, The Journal of biological chemistry.

[11]  M. Kuwano,et al.  A conditional lethal mutation in an Escherichia coli strain with a longer chemical lifetime of messenger RNA. , 1979, Journal of molecular biology.

[12]  G. Shaw,et al.  A conserved AU sequence from the 3′ untranslated region of GM-CSF mRNA mediates selective mRNA degradation , 1986, Cell.

[13]  J. Walker,et al.  Distantly related sequences in the alpha‐ and beta‐subunits of ATP synthase, myosin, kinases and other ATP‐requiring enzymes and a common nucleotide binding fold. , 1982, The EMBO journal.

[14]  P. Lengyel,et al.  Interferon action: binding of viral RNA to the 40-kilodalton 2'-5'-oligoadenylate synthetase in interferon-treated HeLa cells infected with encephalomyocarditis virus , 1991, Journal of virology.

[15]  P. Torrence,et al.  Respective role of each of the purine N-6 amino groups of 5'-O-triphosphoryladenylyl(2'----5')adenylyl(2----5')adenosine in binding to and activation of RNase L. , 1985, The Journal of biological chemistry.

[16]  I. Kerr,et al.  Control of the ppp(a2'p)nA system in HeLa cells. Effects of interferon and virus infection. , 1982, European journal of biochemistry.

[17]  P. R. Sibbald,et al.  The P-loop--a common motif in ATP- and GTP-binding proteins. , 1990, Trends in biochemical sciences.

[18]  G. Stark,et al.  An improved method for purifying 2',5'-oligoadenylate synthetases. , 1984, The Journal of biological chemistry.

[19]  F. Claverie-Martin,et al.  Analysis of the altered mRNA stability (ams) gene from Escherichia coli. Nucleotide sequence, transcriptional analysis, and homology of its product to MRP3, a mitochondrial ribosomal protein from Neurospora crassa. , 1991, The Journal of biological chemistry.

[20]  P. Lengyel,et al.  The interferon system. A bird's eye view of its biochemistry. , 1992, The Journal of biological chemistry.

[21]  J. Dumont,et al.  (2'-5')Oligoadenylate in rat liver: modulation after partial hepatectomy. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[22]  I. Kerr,et al.  Natural occurrence of 2-5A in interferon-treated EMC virus-infected L cells , 1979, Nature.

[23]  I. Kerr,et al.  pppA2'p5'A2'p5'A: an inhibitor of protein synthesis synthesized with an enzyme fraction from interferon-treated cells. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Silverman,et al.  rRNA cleavage as an index of ppp(A2'p)nA activity in interferon-treated encephalomyocarditis virus-infected cells , 1983, Journal of virology.

[25]  I. Kerr,et al.  Analogue inhibitor of 2‐5A action: effect on the interferon‐mediated inhibition of encephalomyocarditis virus replication. , 1985, The EMBO journal.

[26]  M. Kirschner,et al.  Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. , 1977, The Journal of biological chemistry.

[27]  R. Silverman,et al.  Activation of 2-5A-dependent RNase by analogs of 2-5A (5'-O-triphosphoryladenylyl(2'----5')adenylyl(2'----5')adenosine ) using 2',5'-tetraadenylate (core)-cellulose. , 1986, The Journal of biological chemistry.

[28]  A. Lassar,et al.  A conditional lethal mutant of Escherichia coli which affects the processing of ribosomal RNA. , 1978, The Journal of biological chemistry.

[29]  P. Lengyel,et al.  Studies on the role of the 2'-5'-oligoadenylate synthetase-RNase L pathway in beta interferon-mediated inhibition of encephalomyocarditis virus replication , 1988, Journal of virology.

[30]  I. Kerr,et al.  Synthesis and breakdown of pppA2'p5'A2'p5'A and transient inhibiton of protein synthesis in extracts from interferon-treated and control cells. , 1978, European journal of biochemistry.

[31]  I. Kerr,et al.  Synthesis of low molecular weight inhibitor of protein synthesis with enzyme from interferon-treated cells , 1977, Nature.

[32]  I. Kerr,et al.  Distribution of the ppp(A2'p)nA-binding protein and interferon-related enzymes in animals, plants, and lower organisms. , 1982, Biochemical and biophysical research communications.

[33]  I. Kerr,et al.  Ribosomal RNA cleavage, nuclease activation and 2-5A(ppp(A2'p)nA) in interferon-treated cells. , 1981, Nucleic acids research.

[34]  A. Hovanessian,et al.  Anticellular and antiviral effects of pppA(2'p5'A)n. , 1980 .

[35]  P. Sharp,et al.  Molecular cloning of an enhancer binding protein:Isolation by screening of an expression library with a recognition site DNA , 1988, Cell.

[36]  J. Trowsdale,et al.  The ppp(A2'p)nA and protein kinase systems in wild-type and interferon-resistant Daudi cells. , 2005, European journal of biochemistry.

[37]  P. Lengyel,et al.  Interferon action: RNA cleavage pattern of a (2'-5')oligoadenylate--dependent endonuclease. , 1981, Science.

[38]  B. Williams,et al.  Constitutive expression of a 2',5'-oligoadenylate synthetase cDNA results in increased antiviral activity and growth suppression. , 1989, Journal of interferon research.

[39]  R. Silverman,et al.  Photochemical crosslinking in oligonucleotide-protein complexes between a bromine-substituted 2-5A analog and 2-5A-dependent RNase by ultraviolet lamp or laser. , 1990, Analytical biochemistry.

[40]  H. Singh,et al.  Molecular cloning of sequence-specific DNA binding proteins using recognition site probes. , 1989, BioTechniques.

[41]  R. Silverman,et al.  Induction of ppp(A2'p)nA-dependent RNase in murine JLS-V9R cells during growth inhibition. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[42]  S. R. Kushner,et al.  The Ams (altered mRNA stability) protein and ribonuclease E are encoded by the same structural gene of Escherichia coli. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[43]  R. Silverman,et al.  Functional analysis of 2-5A-dependent RNase and 2-5a using 2',5'-oligoadenylate-cellulose. , 1985, Analytical biochemistry.

[44]  B. Williams,et al.  Inhibition of cell-free protein synthesis by pppA2′ p5′ A2′ p5′ A: a novel oligonucleotide synthesized by interferon-treated L cell extracts , 1978, Cell.

[45]  H. V. Rickenberg,et al.  The occurrence of 2'-5' oligoadenylates in Escherichia coli. , 1987, European journal of biochemistry.

[46]  R. Evans,et al.  Zinc fingers: Gilt by association , 1988, Cell.

[47]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[48]  G. Brawerman.  mRNA decay: Finding the right targets , 1989, Cell.

[49]  H. Krisch,et al.  RNase E, an endoribonuclease, has a general role in the chemical decay of Escherichia coli mRNA: evidence that rne and ams are the same genetic locus , 1990, Molecular microbiology.

[50]  R. Silverman,et al.  Regulation of ppp(A2'p)nA-dependent RNase levels during interferon treatment and cell differentiation. , 1985, European journal of biochemistry.

[51]  R. Silverman,et al.  Interferon-induced synthesis of 2-5A-dependent RNase in mouse JLS-V9R cells. , 1983, Virology.

[52]  N. Sonenberg,et al.  A lysine substitution in the ATP-binding site of eucaryotic initiation factor 4A abrogates nucleotide-binding activity , 1989, Molecular and cellular biology.

[53]  P. Lengyel,et al.  Interferon, double-stranded RNA, and RNA degradation: activation of an endonuclease by (2'-5')An. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[54]  D. Storm,et al.  A-type ATP binding consensus sequences are critical for the catalytic activity of the calmodulin-sensitive adenylyl cyclase from Bacillus anthracis. , 1990, The Journal of biological chemistry.

[55]  I. Kerr,et al.  Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells , 1980, Nature.

[56]  R. Slansky.  Theoretical particle physics. , 1981, Science.

[57]  M. Kozak.  Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes , 1986, Cell.

[58]  A. Laurent,et al.  Identification of 69‐kd and 100‐kd forms of 2‐5A synthetase in interferon‐treated human cells by specific monoclonal antibodies. , 1987, The EMBO journal.

[59]  J. Skehel,et al.  Interferon action—sequence specificity of the ppp(A2′p)nA-dependent ribonuclease , 1981, Nature.