Article
Mutational analysis of bacteriophage T4 RNA ligase 1. Different functional groups are required for the nucleotidyl transfer and phosphodiester bond formation steps of the ligation reaction.
The Journal of biological chemistry - 8 Aug 2003
Wang Li Kai, Ho C Kiong, Pei Yi, Shuman Stewart
Abstract excerpt
T4 RNA ligase 1 (Rnl1) exemplifies an ATP-dependent RNA ligase family that includes fungal tRNA ligase (Trl1) and a putative baculovirus RNA ligase. Rnl1 acts via a covalent enzyme-AMP intermediate generated by attack of Lys-99 N zeta on the alpha phosphorus of ATP. Mutation of Lys-99 abolishes ligase activity. Here we tested the effects of alanine mutations at 19 conserved positions in Rnl1 and thereby...
Topics
- Alanine
- Amino Acid Motifs
- Amino Acid Sequence
- Arginine
- Bacteriophage T4
- Binding Sites
- DNA Mutational Analysis
- Electrophoresis, Agar Gel
- Lysine
- Molecular Sequence Data
- Mutation
- Phenylalanine
- Protein Structure, Tertiary