scholarly journals Characterization of a Trifunctional Mimivirus mRNA Capping Enzyme and Crystal Structure of the RNA Triphosphatase Domain

Structure ◽  
2008 ◽  
Vol 16 (4) ◽  
pp. 501-512 ◽  
Author(s):  
Delphine Benarroch ◽  
Paul Smith ◽  
Stewart Shuman
Keyword(s):  
Crystal Structure ◽  
Mrna Capping ◽  
Rna Triphosphatase ◽  
Capping Enzyme

scholarly journals Crystal Structure of Vaccinia Virus mRNA Capping Enzyme Provides Insights into the Mechanism and Evolution of the Capping Apparatus

Structure ◽  
2014 ◽  
Vol 22 (3) ◽  
pp. 452-465 ◽  
Author(s):  
Otto J.P. Kyrieleis ◽  
Jonathan Chang ◽  
Marcos de la Peña ◽  
Stewart Shuman ◽  
Stephen Cusack
Keyword(s):  
Crystal Structure ◽  
Vaccinia Virus ◽  
Mrna Capping ◽  
Capping Enzyme

Purification and characterization of the mRNA capping enzyme GTP:RNA guanylyltransferase from wheat germ

Biochemistry ◽  
1982 ◽  
Vol 21 (2) ◽  
pp. 327-333 ◽  
Author(s):  
Jerry M. Keith ◽  
Sundararajan Venkatesan ◽  
Alan Gershowitz ◽  
Bernard Moss
Keyword(s):  
Wheat Germ ◽  
Purification And Characterization ◽  
Mrna Capping ◽  
Capping Enzyme

Isolation and Characterization of the Yeast mRNA Capping Enzyme β Subunit Gene Encoding RNA 5′-Triphosphatase, Which Is Essential for Cell Viability

1997 ◽  
Vol 239 (1) ◽  
pp. 116-122 ◽  
Author(s):  
Toshihiko Tsukamoto ◽  
Yoshio Shibagaki ◽  
Shinobu Imajoh-Ohmi ◽  
Teruko Murakoshi ◽  
Masako Suzuki ◽  
...  
Keyword(s):  
Cell Viability ◽  
Β Subunit ◽  
Subunit Gene ◽  
Gene Encoding ◽  
Isolation And Characterization ◽  
Mrna Capping ◽  
Capping Enzyme

Cloning and Characterization of Two Human cDNAs Encoding the mRNA Capping Enzyme

1998 ◽  
Vol 243 (1) ◽  
pp. 101-108 ◽  
Author(s):  
Toshihiko Tsukamoto ◽  
Yoshio Shibagaki ◽  
Teruko Murakoshi ◽  
Masako Suzuki ◽  
Akiko Nakamura ◽  
...  
Keyword(s):  
Mrna Capping ◽  
Capping Enzyme

scholarly journals The Essential Role for the RNA Triphosphatase Cet1p in Nuclear Import of the mRNA Capping Enzyme Cet1p-Ceg1p Complex of Saccharomyces cerevisiae

PLoS ONE ◽  
2013 ◽  
Vol 8 (10) ◽  
pp. e78000
Author(s):  
Naoki Takizawa ◽  
Toshinobu Fujiwara ◽  
Manabu Yamasaki ◽  
Ayako Saito ◽  
Akira Fukao ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nuclear Import ◽  
Essential Role ◽  
Mrna Capping ◽  
Rna Triphosphatase ◽  
Capping Enzyme

scholarly journals A Yeast-Based Genetic System for Functional Analysis of Viral mRNA Capping Enzymes

2000 ◽  
Vol 74 (12) ◽  
pp. 5486-5494 ◽  
Author(s):  
C. Kiong Ho ◽  
Alexandra Martins ◽  
Stewart Shuman
Keyword(s):  
Fusion Protein ◽  
Vaccinia Virus ◽  
Rna Polymerase Ii ◽  
Elongation Complex ◽  
Mrna Capping ◽  
Rna Triphosphatase ◽  
Capping Enzyme ◽  
Capping Enzymes

ABSTRACT Virus-encoded mRNA capping enzymes are attractive targets for antiviral therapy, but functional studies have been limited by the lack of genetically tractable in vivo systems that focus exclusively on the RNA-processing activities of the viral proteins. Here we have developed such a system by engineering a viral capping enzyme—vaccinia virus D1(1-545)p, an RNA triphosphatase and RNA guanylyltransferase—to function in the budding yeast Saccharomyces cerevisiae in lieu of the endogenous fungal triphosphatase (Cet1p) and guanylyltransferase (Ceg1p). This was accomplished by fusion of D1(1-545)p to the C-terminal guanylyltransferase domain of mammalian capping enzyme, Mce1(211-597)p, which serves as a vehicle to target the viral capping enzyme to the RNA polymerase II elongation complex. An inactivating mutation (K294A) of the mammalian guanylyltransferase active site in the fusion protein had no impact on genetic complementation of cet1Δceg1Δ cells, thus proving that (i) the viral guanylyltransferase was active in vivo and (ii) the mammalian domain can serve purely as a chaperone to direct other proteins to the transcription complex. Alanine scanning had identified five amino acids of vaccinia virus capping enzyme—Glu37, Glu39, Arg77, Glu192, and Glu194—that are essential for γ phosphate cleavage in vitro. Here we show that the introduction of mutation E37A, R77A, or E192A into the fusion protein abrogates RNA triphosphatase function in vivo. The essential residues are located within three motifs that define a family of viral and fungal metal-dependent phosphohydrolases with a distinctive capacity to hydrolyze nucleoside triphosphates to nucleoside diphosphates in the presence of manganese or cobalt. The acidic residues Glu37, Glu39, and Glu192 likely comprise the metal-binding site of vaccinia virus triphosphatase, insofar as their replacement by glutamine abolishes the RNA triphosphatase and ATPase activities.

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