Structure-function analysis of T4 RNA ligase 2 Journal Article
| Authors: | Yin, S.; Ho, C. K.; Shuman, S. |
| Article Title: | Structure-function analysis of T4 RNA ligase 2 |
| Abstract: | Bacteriophage T4 RNA ligase 2 (Rnl2) exemplifies a polynucleotide ligase family that includes the trypanosome RNA-editing ligases and putative RNA ligases encoded by eukaryotic viruses and archaea. Here we analyzed 12 individual amino acids of Rnl2 that were identified by alanine scanning as essential for strand joining. We determined structure-activity relationships via conservative substitutions and examined mutational effects on the isolated steps of ligase adenylylation and phosphodiester bond formation. The essential residues of Rnl2 are located within conserved motifs that define a superfamily of nucleotidyl transferases that act via enzyme-(lysyl-N)-NMP intermediates. Our mutagenesis results underscore a shared active site architecture in Rnl2-like ligases, DNA ligases, and mRNA capping enzymes. They also highlight two essential signature residues, Glu34 and Asn40, that flank the active site lysine nucleophile (Lys35) and are unique to the Rnl2-like ligase family. |
| Keywords: | unclassified drug; genetics; mutation; dose response; nonhuman; molecular genetics; metabolism; amino acid substitution; dose-response relationship, drug; trypanosoma; structure activity relation; structure-activity relationship; mutational analysis; physiology; rna; chemistry; amino acid sequence; molecular sequence data; sequence homology, amino acid; messenger rna; eukaryota; recombinant proteins; recombinant protein; binding site; mutagenesis, site-directed; binding sites; molecular structure; alanine; adenosine triphosphate; conformation; nucleic acid conformation; sequence homology; polydeoxyribonucleotide synthase; site directed mutagenesis; biochemistry; glutamic acid; mutagenesis; enzymes; lysine; archaebacterium; rna ligase (atp); archaea; asparagine; eukaryote; esters; chemical bond; virus protein; viral proteins; amino acid analysis; adenylation; nucleotidyltransferase; rna capping; viruses; rna ligase; polynucleotide ligases; bacteriophage t4; rna ligase 2; priority journal; article; unidentified bacteriophage; strand joining; bacteriophage t4 rna ligase 2 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 278 |
| Issue: | 20 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2003-05-16 |
| Start Page: | 17601 |
| End Page: | 17608 |
| Language: | English |
| DOI: | 10.1074/jbc.M300817200 |
| PUBMED: | 12611899 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 12 September 2014 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work