JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Saha, N.
Right arrow Articles by Shuman, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Saha, N.
Right arrow Articles by Shuman, S.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

J Biol Chem, Vol. 274, Issue 23, 16553-16562, June 4, 1999

Characterization of Human, Schizosaccharomyces pombe, and Candida albicans mRNA Cap Methyltransferases and Complete Replacement of the Yeast Capping Apparatus by Mammalian Enzymes

Nayanendu SahaDagger , Beate Schwer§, and Stewart ShumanDagger

From the Dagger  Molecular Biology Program, Sloan-Kettering Institute and the § Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, New York 10021

Human and fission yeast cDNAs encoding mRNA (guanine-N7) methyltransferase were identified based on similarity of the human (Hcm1p; 476 amino acids) and Schizosaccharomyces pombe (Pcm1p; 389 amino acids) polypeptides to the cap methyltransferase of Saccharomyces cerevisiae (Abd1p). Expression of PCM1 or HCM1 in S. cerevisiae complemented the lethal phenotype resulting from deletion of the ABD1 gene, as did expression of the NH2-terminal deletion mutants PCM1(94-389) and HCM1(121-476). The CCM1 gene encoding Candida albicans cap methyltransferase (Ccm1p; 474 amino acids) was isolated from a C. albicans genomic library by selection for complementation of the conditional growth phenotype of S. cerevisiae abd1-ts mutants. Human cap methyltransferase was expressed in bacteria, purified, and characterized. Recombinant Hcm1p catalyzed quantitative S-adenosylmethionine-dependent conversion of GpppA-capped poly(A) to m7GpppA-capped poly(A). We identified by alanine-scanning mutagenesis eight amino acids (Asp-203, Gly-207, Asp-211, Asp-227, Arg-239, Tyr-289, Phe-291, and Phe-354) that are essential for human cap methyltransferase function in vivo. All eight residues are conserved in other cellular cap methyltransferases. Five of the mutant human proteins (D203A, R239A, Y289A, F291A, and F354A) were expressed in bacteria and found to be defective in cap methylation in vitro. Concordance of mutational effects on Hcm1p, Abd1p, and vaccinia capping enzyme underscores a conserved structural basis for cap methylation in DNA viruses, yeast, and metazoans. This is in contrast to the structural and mechanistic divergence of the RNA triphosphatase components of the yeast and metazoan capping systems. Nevertheless, we demonstrate that the entire three-component yeast capping apparatus, consisting of RNA 5'-triphosphatase (Cet1p), RNA guanylyltransferase (Ceg1p), and Abd1p could be replaced in vivo by the two-component mammalian apparatus consisting of a bifunctional triphosphatase-guanylyltransferase Mce1p and the methyltransferase Hcm1(121-476)p. Isogenic yeast strains with fungal versus mammalian capping systems should facilitate rational screens for antifungal drugs that target cap formation in vivo.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 RNAHome page
S. Zheng and S. Shuman
Structure-function analysis of vaccinia virus mRNA cap (guanine-N7) methyltransferase
RNA, April 1, 2008; 14(4): 696 - 705.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Zheng, S. Shuman, and B. Schwer
Sinefungin resistance of Saccharomyces cerevisiae arising from sam3 mutations that inactivate the AdoMet transporter or from increased expression of AdoMet synthase plus mRNA cap guanine-N7 methyltransferase
Nucleic Acids Res., November 29, 2007; 35(20): 6895 - 6903.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Takagi, S. Sindkar, D. Ekonomidis, M. P. Hall, and C. K. Ho
Trypanosoma brucei Encodes a Bifunctional Capping Enzyme Essential for Cap 4 Formation on the Spliced Leader RNA
J. Biol. Chem., June 1, 2007; 282(22): 15995 - 16005.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
S. Hausmann, A. Ramirez, S. Schneider, B. Schwer, and S. Shuman
Biochemical and genetic analysis of RNA cap guanine-N2 methyltransferases from Giardia lamblia and Schizosaccharomyces pombe
Nucleic Acids Res., March 12, 2007; 35(5): 1411 - 1420.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
F. Peyrane, B. Selisko, E. Decroly, J. J. Vasseur, D. Benarroch, B. Canard, and K. Alvarez
High-yield production of short GpppA- and 7MeGpppA-capped RNAs and HPLC-monitoring of methyltransfer reactions at the guanine-N7 and adenosine-2'O positions
Nucleic Acids Res., February 28, 2007; 35(4): e26 - e26.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Zheng, S. Hausmann, Q. Liu, A. Ghosh, B. Schwer, C. D. Lima, and S. Shuman
Mutational Analysis of Encephalitozoon cuniculi mRNA Cap (Guanine-N7) Methyltransferase, Structure of the Enzyme Bound to Sinefungin, and Evidence That Cap Methyltransferase Is the Target of Sinefungin's Antifungal Activity
J. Biol. Chem., November 24, 2006; 281(47): 35904 - 35913.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
B. Schwer, S. Hausmann, S. Schneider, and S. Shuman
Poxvirus mRNA Cap Methyltransferase: BYPASS OF THE REQUIREMENT FOR THE STIMULATORY SUBUNIT BY MUTATIONS IN THE CATALYTIC SUBUNIT AND EVIDENCE FOR INTERSUBUNIT ALLOSTERY
J. Biol. Chem., July 14, 2006; 281(28): 18953 - 18960.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
M. P. HALL and C. K. HO
Characterization of a Trypanosoma brucei RNA cap (guanine N-7) methyltransferase
RNA, March 1, 2006; 12(3): 488 - 497.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
Y. Pei, H. Du, J. Singer, C. St. Amour, S. Granitto, S. Shuman, and R. P. Fisher
Cyclin-Dependent Kinase 9 (Cdk9) of Fission Yeast Is Activated by the CDK-Activating Kinase Csk1, Overlaps Functionally with the TFIIH-Associated Kinase Mcs6, and Associates with the mRNA Cap Methyltransferase Pcm1 In Vivo
Mol. Cell. Biol., February 1, 2006; 26(3): 777 - 788.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
G. L. Chrebet, D. Wisniewski, A. L. Perkins, Q. Deng, M. B. Kurtz, A. Marcy, and S. A. Parent
Cell-Based Assays to Detect Inhibitors of Fungal mRNA Capping Enzymes and Characterization of Sinefungin as a Cap Methyltransferase Inhibitor
J Biomol Screen, June 1, 2005; 10(4): 355 - 364.
[Abstract] [PDF]

Home page
 J. Biol. Chem.Home page
S. Hausmann, S. Zheng, C. Fabrega, S. W. Schneller, C. D. Lima, and S. Shuman
Encephalitozoon cuniculi mRNA Cap (Guanine N-7) Methyltransferase: METHYL ACCEPTOR SPECIFICITY, INHIBITION BY S-ADENOSYLMETHIONINE ANALOGS, AND STRUCTURE-GUIDED MUTATIONAL ANALYSIS
J. Biol. Chem., May 27, 2005; 280(21): 20404 - 20412.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
B. Shafer, C. Chu, and A. J. Shatkin
Human mRNA Cap Methyltransferase: Alternative Nuclear Localization Signal Motifs Ensure Nuclear Localization Required for Viability
Mol. Cell. Biol., April 1, 2005; 25(7): 2644 - 2649.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Hausmann, M. A. Altura, M. Witmer, S. M. Singer, H. G. Elmendorf, and S. Shuman
Yeast-like mRNA Capping Apparatus in Giardia lamblia
J. Biol. Chem., April 1, 2005; 280(13): 12077 - 12086.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
N. Saha, S. Shuman, and B. Schwer
Yeast-Based Genetic System for Functional Analysis of Poxvirus mRNA Cap Methyltransferase
J. Virol., July 1, 2003; 77(13): 7300 - 7307.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Srinivasan, F. Piano, and A. J. Shatkin
mRNA Capping Enzyme Requirement for Caenorhabditis elegans Viability
J. Biol. Chem., April 11, 2003; 278(16): 14168 - 14173.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
D. S. Dunyak, D. S. Everdeen, J. G. Albanese, and C. L. Quinn
Deletion of Individual mRNA Capping Genes Is Unexpectedly Not Lethal to Candida albicans and Results in Modified mRNA Cap Structures
Eukaryot. Cell, December 1, 2002; 1(6): 1010 - 1020.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. Pei, B. Schwer, S. Hausmann, and S. Shuman
Characterization of Schizosaccharomyces pombe RNA triphosphatase
Nucleic Acids Res., January 15, 2001; 29(2): 387 - 396.
[Abstract] [Full Text] [PDF]

Home page
 Genes Dev.Home page
Y. Wen and A. J. Shatkin
Cap methyltransferase selective binding and methylation of GpppG-RNA are stimulated by importin-{alpha}
Genes & Dev., December 1, 2000; 14(23): 2944 - 2949.
[Abstract] [Full Text]

Home page
 GeneticsHome page
B. Schwer, N. Saha, X. Mao, H.-W. Chen, and S. Shuman
Structure-Function Analysis of Yeast mRNA Cap Methyltransferase and High-Copy Suppression of Conditional Mutants by AdoMet Synthase and the Ubiquitin Conjugating Enzyme Cdc34p
Genetics, August 1, 2000; 155(4): 1561 - 1576.
[Abstract] [Full Text]

Home page
 J. Virol.Home page
C. K. Ho, A. Martins, and S. Shuman
A Yeast-Based Genetic System for Functional Analysis of Viral mRNA Capping Enzymes
J. Virol., June 15, 2000; 74(12): 5486 - 5494.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
Y. Pei, C. K. Ho, B. Schwer, and S. Shuman
Mutational Analyses of Yeast RNA Triphosphatases Highlight a Common Mechanism of Metal-dependent NTP Hydrolysis and a Means of Targeting Enzymes to Pre-mRNAs in Vivo by Fusion to the Guanylyltransferase Component of the Capping Apparatus
J. Biol. Chem., October 8, 1999; 274(41): 28865 - 28874.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
B. Schwer, K. Lehman, N. Saha, and S. Shuman
Characterization of the mRNA Capping Apparatus of Candida albicans
J. Biol. Chem., January 12, 2001; 276(3): 1857 - 1864.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Hausmann, C. P. Vivares, and S. Shuman
Characterization of the mRNA Capping Apparatus of the Microsporidian Parasite Encephalitozoon cuniculi
J. Biol. Chem., January 4, 2002; 277(1): 96 - 103.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. Vasiljeva, A. Merits, P. Auvinen, and L. Kaariainen
Identification of a Novel Function of the Alphavirus Capping Apparatus. RNA 5'-TRIPHOSPHATASE ACTIVITY OF Nsp2
J. Biol. Chem., June 2, 2000; 275(23): 17281 - 17287.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 1999 by the American Society for Biochemistry and Molecular Biology.