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Biochemical and genetic analysis of RNA cap guanine-N2 methyltransferases from Giardia lamblia and Schizosaccharomyces pombe by S. Hausmann; A. Ramirez; S. Schneider; B. Schwer; S. Shuman is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Biochemical and genetic analysis of RNA cap guanine-N2 methyltransferases from Giardia lamblia and Schizosaccharomyces pombe about?

RNA cap guanine-N2 methyltransferases such as __Schizosaccharomyces pombe__ Tgs1 and __Giardia lamblia__ Tgs2 catalyze methylation of the exocyclic N2 amine of 7-methylguanosine. Here we performed a mutational analysis of __Giardia__ Tgs2, entailing an alanine scan of 17 residues within the minimal active domain. Alanine substitutions at Phe18, Thr40, Asp76, Asn103 and Asp140 reduced methyltransferase specific activity to <3% of wild-type Tgs2, thereby defining these residues as essential. Alanines at Pro142, Tyr148 and Pro185 reduced activity to 7–12% of wild-type. Structure–activity relationships at Phe18, Thr40, Asp76, Asn103, Asp140 and Tyr148, and at three other essential residues defined previously (Asp68, Glu91 and Trp143) were gleaned by testing the effects of 18 conservative substitutions. Our results engender a provisional map of the Tgs2 active site, which we discuss in light of crystal structures of related methyltransferases. A genetic analysis of __S. pombe__ Tgs1 showed that it is nonessential. An __S. pombe tgs1__Δ strain grows normally, notwithstanding the absence of 2,2,7-trimethylguanosine caps on its U1, U2, U4 and U5 snRNAs. However, we find that __S. pombe__ r

Who reads Biochemical and genetic analysis of RNA cap guanine-N2 methyltransferases from Giardia lamblia and Schizosaccharomyces pombe?

Author

S. Hausmann; A. Ramirez; S. Schneider; B. Schwer; S. Shuman

Publisher

Oxford University Press; London: Information Retrieval; Oxford: Oxford University Press; Oxford University Press (OUP) (ISSN 0305-1048)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)