Crystal structure of eukaryotic DNA ligase-adenylate illuminates the mechanism of nick sensing and strand joining Journal Article
| Authors: | Odell, M.; Sriskanda, V.; Shuman, S.; Nikolov, D. B. |
| Article Title: | Crystal structure of eukaryotic DNA ligase-adenylate illuminates the mechanism of nick sensing and strand joining |
| Abstract: | Chlorella virus DNA ligase is the smallest eukaryotic ATP-dependent ligase known; it has an intrinsic nick-sensing function and suffices for yeast cell growth. Here we report the 2.0 Å crystal structure of the covalent ligase-AMP reaction intermediate. The conformation of the adenosine nucleoside and contacts between the enzyme and the ribose sugar have undergone a significant change compared to complexes of T7 ligase with ATP or mRNA capping enzymes with GTP. The conformational switch allows the 3' OH of AMP to coordinate directly the 5' PO(4) of the nick. The structure explains why nick sensing is restricted to adenylated ligase and why the 5' phosphate is required for DNA binding. We identify a metal binding site on ligase-adenylate and propose a mechanism of nick recognition and catalysis supported by mutational data. |
| Keywords: | mutation; dna-binding proteins; nonhuman; cell growth; dna; amino acid sequence; molecular sequence data; messenger rna; adenosine; sequence alignment; recombination, genetic; base sequence; crystal structure; models, molecular; crystallography, x-ray; protein structure, tertiary; binding sites; molecular interaction; conformational transition; catalysis; adenosine triphosphate; dna binding; structure analysis; polydeoxyribonucleotide synthase; guanosine triphosphate; phosphates; yeast cell; dna nick translation; arginine; eukaryote; metals; chlorella virus; dna ligases; viral proteins; enzyme conformation; adenosine phosphate; cations, divalent; nucleotidyltransferases; dna strand; adenosine monophosphate; eukaryotic cells; electrostatics; article; bacteriophage t7 |
| Journal Title: | Molecular Cell |
| Volume: | 6 |
| Issue: | 5 |
| ISSN: | 1097-2765 |
| Publisher: | Cell Press |
| Date Published: | 2000-11-01 |
| Start Page: | 1183 |
| End Page: | 1193 |
| Language: | English |
| PUBMED: | 11106756 |
| PROVIDER: | scopus |
| DOI: | 10.1016/S1097-2765(00)00115-5 |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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