Conversion of a DNA ligase into an RNA capping enzyme.

In eukaryotes, newly synthesised mRNA is 'capped' by the addition of GMP to the 5" end by RNA capping enzymes. Recent structural studies have shown that RNA capping enzymes and DNA ligases have similar protein folds, suggesting a conserved catalytic mechanism. To explore these similarities we have produced a chimeric enzyme comprising the N-terminal domain 1 of a DNA ligase fused to the C-terminal domain 2 of a mRNA capping enzyme. This report shows that this hybrid enzyme retains adenylation activity, characteristic of DNA ligases but, remarkably, the chimera has ATP-dependent mRNA capping activity. This is the first observation of ATP-dependent RNA capping. These results suggest that nucleotidyltransferases may have evolved from a common ancestral gene.

[1]  D. Wigley,et al.  Functional domains of an ATP-dependent DNA ligase. , 1999, Journal of molecular biology.

[2]  S. Shuman,et al.  Mutational analysis of Chlorella virus DNA ligase: catalytic roles of domain I and motif VI. , 1998, Nucleic acids research.

[3]  S. Shuman,et al.  Chlorella virus DNA ligase: nick recognition and mutational analysis. , 1998, Nucleic acids research.

[4]  C. Ho,et al.  Phylogeny of mRNA capping enzymes. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Stewart Shuman,et al.  X-Ray Crystallography Reveals a Large Conformational Change during Guanyl Transfer by mRNA Capping Enzymes , 1997, Cell.

[6]  D. Suck.  Common fold, common function, common origin? , 1997, Nature Structural Biology.

[7]  J. V. Van Etten,et al.  Expression and characterization of an RNA capping enzyme encoded by Chlorella virus PBCV-1 , 1996, Journal of virology.

[8]  D. Wigley,et al.  Characterization of proteolytic fragments of bacteriophage T7 DNA ligase. , 1996, Nucleic acids research.

[9]  D. Wigley,et al.  Crystal Structure of an ATP-Dependent DNA Ligase from Bacteriophage T7 , 1996, Cell.

[10]  D. Wigley,et al.  Bacteriophage T7 DNA Ligase , 1996, The Journal of Biological Chemistry.

[11]  S. Shuman,et al.  Mutational analysis of mRNA capping enzyme identifies amino acids involved in GTP binding, enzyme-guanylate formation, and GMP transfer to RNA , 1995, Molecular and cellular biology.

[12]  S. Shuman,et al.  Mutational analysis of vaccinia DNA ligase defines residues essential for covalent catalysis. , 1995, Virology.

[13]  S. Shuman,et al.  RNA capping enzyme and DNA ligase: a superfamily of covalent nucleotidyl transferases , 1995, Molecular microbiology.

[14]  S. Shuman.  Capping enzyme in eukaryotic mRNA synthesis. , 1995, Progress in nucleic acid research and molecular biology.

[15]  S. Shuman,et al.  Covalent catalysis in nucleotidyl transfer reactions: essential motifs in Saccharomyces cerevisiae RNA capping enzyme are conserved in Schizosaccharomyces pombe and viral capping enzymes and among polynucleotide ligases. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S. Buratowski,et al.  Active site of the mRNA-capping enzyme guanylyltransferase from Saccharomyces cerevisiae: similarity to the nucleotidyl attachment motif of DNA and RNA ligases. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[17]  T. Lindahl,et al.  In vitro mutagenesis and functional expression in Escherichia coli of a cDNA encoding the catalytic domain of human DNA ligase I. , 1991, Nucleic acids research.

[18]  S. Heaphy,et al.  Effect of single amino acid changes in the region of the adenylylation site of T4 RNA ligase. , 1987, Biochemistry.

[19]  S. Shuman.  RNA capping by HeLa cell RNA guanylyltransferase. Characterization of a covalent protein-guanylate intermediate. , 1982, The Journal of biological chemistry.

[20]  B. Moss,et al.  Eukaryotic mRNA capping enzyme-guanylate covalent intermediate. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Hurwitz,et al.  Mechanism of mRNA capping by vaccinia virus guanylyltransferase: characterization of an enzyme--guanylate intermediate. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Surks,et al.  Purification and characterization of a GTP-pyrophosphate exchange activity from vaccinia virions. Association of the GTP-pyrophosphate exchange activity with vaccinia mRNA guanylyltransferase . RNA (guanine-7-)methyltransferase complex (capping enzyme). , 1980, The Journal of biological chemistry.

[23]  I. R. Lehnman,et al.  DNA Ligase: Structure, Mechanism, and Function , 1974, Science.

[24]  V. Georgiev.  Virology , 1955, Nature.