Characterization of a baculovirus enzyme with RNA ligase, polynucleotide 5′-kinase, and polynucleotide 3′-phosphatase activities Journal Article
| Authors: | Martins, A.; Shuman, S. |
| Article Title: | Characterization of a baculovirus enzyme with RNA ligase, polynucleotide 5′-kinase, and polynucleotide 3′-phosphatase activities |
| Abstract: | The end-healing and end-sealing steps of the phage T4-induced RNA restriction-repair pathway are performed by two separate enzymes, a bifunctional polynucleotide 5′-kinase/3′-phosphatase and an ATP-dependent RNA ligase. Here we show that a single trifunctional baculovirus enzyme, RNA ligase 1 (Rnl1), catalyzes the identical set of RNA repair reactions. Three enzymatic activities of baculovirus Rnl1 are organized in a modular fashion within a 694-amino acid polypeptide consisting of an autonomous N-terminal RNA-specific ligase domain, Rnl1-(1-385), and a C-terminal kinase-phosphatase domain, Rnl1-(394-694). The ligase domain is itself composed of two functional units. The N-terminal module Rnl1-(1-270) contains essential nucleotidyltransferase motifs I, IV, and V and suffices for both enzyme adenylylation (step 1 of the ligation pathway) and phosphodiester bond formation at a preactivated RNA-adenylate end (step 3). The downstream module extending to residue 385 is required for ligation of a phosphorylated RNA substrate, suggesting that it is involved specifically in the second step of the end-joining pathway, the transfer of AMP from the ligase to the 5′-PO4 end to form RNA-adenylate. The end-healing domain Rnl1-(394-694) consists of a proximal 5′-kinase module with an essential P-loop motif (404GSGKS408) and a distal 3′-phosphatase module with an essential acylphosphatase motif ( 560DLDGT564). Our findings have implications for the evolution of RNA repair systems and their potential roles in virus-host dynamics. |
| Keywords: | unclassified drug; nonhuman; protein domain; phosphatase; carboxy terminal sequence; enzyme activity; rna; cloning, molecular; amino acid sequence; amino terminal sequence; enzyme analysis; dna viruses; sequence alignment; amino acid; protein structure, tertiary; catalysis; adenosine triphosphate; multienzyme complexes; rna processing; biochemistry; phosphate; polynucleotide 5' hydroxyl kinase; rna ligase (atp); hydrogen-ion concentration; metals; viral proteins; adenosine phosphate; adenylation; nucleotidyltransferase; polypeptide; rna repair; virus cell interaction; rna ligase; baculovirus; baculoviridae; bacteriophage t4; adenosinetriphosphate; unidentified baculovirus; phosphotransferases (phosphate group acceptor); polyribonucleotide nucleotidyltransferase; priority journal; article; enzymatic activities; enzyme adenylylation; aspartylleucylaspartylglycylthreonine; glycylserylglycyllysylserine; polynucleotide 3' phosphatase; polynucleotide adenylyltransferase; rna ligase 1 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 279 |
| Issue: | 18 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2004-04-30 |
| Start Page: | 18220 |
| End Page: | 18231 |
| Language: | English |
| DOI: | 10.1074/jbc.M313386200 |
| PROVIDER: | scopus |
| PUBMED: | 14747466 |
| DOI/URL: | |
| Notes: | J. Biol. Chem. -- Cited By (since 1996):31 -- Export Date: 16 June 2014 -- CODEN: JBCHA -- Source: Scopus |
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