Structure-function analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase

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Mutational analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase reveals essential contributions of the N-terminal peptide that closes over the active site.

RNA guanine-N7 methyltransferase catalyzes the third step of eukaryal mRNA capping, the transfer of a methyl group from AdoMet to GpppRNA to form m(7)GpppRNA. Mutational and crystallographic analyses of cellular and poxvirus cap methyltransferases have yielded a coherent picture of a conserved active site and determinants of substrate specificity. Models of the Michaelis complex suggest a direc...

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Structural insights into the mechanism and evolution of the vaccinia virus mRNA cap N7 methyl-transferase.

The vaccinia virus mRNA capping enzyme is a multifunctional heterodimeric protein associated with the viral polymerase that both catalyses the three steps of mRNA capping and regulates gene transcription. The structure of a subcomplex comprising the C-terminal N7-methyl-transferase (MT) domain of the large D1 subunit, the stimulatory D12 subunit and bound S-adenosyl-homocysteine (AdoHcy) has be...

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mRNA ( nucleoside - 2 ’ - ) - methyltransferase from Vaccinia Virus

An S-adenosyl-L-methionine:mRNA(nucleoside-2’-)methyltransferase, one of at least three activities required for the 5’-terminal modification of mRNA, has been purified from vaccinia virus particles. Employing brome mosaic virus RNA ending in m7G(5’)pppGas substrate, a simple DEAE-cellulose filter assay measuring the incorporation of methyl groups from S-adenosyl[methyL3H]methionine to position ...

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Purification of mRNA guanylyltransferase and mRNA (guanine-7-) methyltransferase from vaccinia virions.

The sequences m7G(5')pppGm-and m7G(5')pppAm-are located at the 5' termini of vaccinia mRNAs. Two novel enzymatic activities have been purified from vaccinia virus cores which modify the 5' terminus of unmethylated mRNA. One activity transfers GMP from GTP to mRNA and is designated a GTP: mRNA guanylyltransferase. The second activity transfers a methyl group from S-adenosylmethionine to position...

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Structure-function analysis of the triphosphatase component of vaccinia virus mRNA capping enzyme.

The N-terminal 60 kDa (amino acids 1 to 545) of the D1 subunit of vaccinia virus mRNA capping enzyme is an autonomous bifunctional domain with triphosphatase and guanylyltransferase activities. We previously described two alanine cluster mutations, R77 to A (R77A)-K79A and E192A-E194A, which selectively inactivated the triphosphatase component. Here, we characterize the activities of 11 single ...

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ژورنال

عنوان ژورنال: RNA

سال: 2008

ISSN: 1355-8382

DOI: 10.1261/rna.928208