Divergent subunit interactions among fungal mRNA 5'-capping machineries.
The Saccharomyces cerevisiae mRNA capping enzyme consists of two subunits: an RNA 5'-triphosphatase (RTPase) and GTP::mRNA guanylyltransferase (GTase). The GTase subunit (Ceg1) binds to the phosphorylated carboxyl-terminal domain of the largest subunit (CTD-P) of RNA polymerase II (pol II), coupling capping with transcription. Ceg1 bound to the CTD-P is inactive unless allosterically activated by interaction with the RTPase subunit (Cet1). For purposes of comparison, we characterize here the related GTases and RTPases from the yeasts Schizosaccharomyces pombe and Candida albicans. Surprisingly, the S. pombe capping enzyme subunits do not interact with each other. Both can independently interact with CTD-P of pol II, and the GTase is not repressed by CTD-P binding. The S. pombe RTPase gene (pct1+) is essential for viability. Pct1 can replace the S. cerevisiae RTPase when GTase activity is supplied by the S. pombe or mouse enzymes but not by the S. cerevisiae GTase. The C. albicans capping enzyme subunits do interact with each other. However, this interaction is not essential in vivo. Our results reveal an unexpected diversity among the fungal capping machineries.
Takagi
Toshimitsu
T
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 021151, USA.
Cho
Eun-Jung
EJ
Janoo
Rozmin T K
RT
Polodny
Vladimir
V
Takase
Yasutaka
Y
Keogh
Michael C
MC
Woo
Sue-Ann
SA
Fresco-Cohen
Lucille D
LD
Hoffman
Charles S
CS
Buratowski
Stephen
S
D017766:Acid Anhydride Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics; D004257:DNA Polymerase II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004271:DNA, Fungal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005658:Fungi / Q000201:enzymology* / Q000235:genetics; D005800:Genes, Fungal; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D021122:Protein Subunits; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D000098556:mRNA Guanylyltransferases
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D021122:Protein Subunits; D029702:Schizosaccharomyces pombe Proteins; C087021:res2 protein, S pombe; D009713:Nucleotidyltransferases; D000098556:mRNA Guanylyltransferases; D004257:DNA Polymerase II; D017766:Acid Anhydride Hydrolases; C023090:RNA triphosphatase