Characterization of a novel eukaryal nick-sealing RNA ligase from Naegleria gruberi Journal Article
| Authors: | Unciuleac, M. C.; Shuman, S. |
| Article Title: | Characterization of a novel eukaryal nick-sealing RNA ligase from Naegleria gruberi |
| Abstract: | The proteome of the amoebo-flagellate protozoan Naegleria gruberi is rich in candidate RNA repair enzymes, including 15 putative RNA ligases, one of which, NgrRnl, is a eukaryal homolog of Deinococcus radiodurans RNA ligase, DraRnl. Here we report that purified recombinant NgrRnl seals nicked 3′-OH/5′-PO<inf>4</inf> duplexes in which the 3′-OH strand is RNA. It does so via the "classic" ligase pathway, entailing reaction with ATP to form a covalent NgrRnl-AMP intermediate, transfer of AMP to the nick 5′-PO<inf>4</inf>, and attack of the RNA 3′-OH on the adenylylated nick to form a 3′-5′ phosphodiester. Unlike members of the four known families of ATP-dependent RNA ligases, NgrRnl lacks a carboxy-terminal appendage to its nucleotidyltransferase domain. Instead, it contains a defining amino-terminal domain that we show is important for 3′-OH/5′-PO<inf>4</inf> nick-sealing and ligase adenylylation, but dispensable for phosphodiester synthesis at a preadenylylated nick. We propose that NgrRnl, DraRnl, and their homologs from diverse bacteria, viruses, and unicellular eukarya comprise a new "Rnl5 family" of nick-sealing ligases with a signature domain organization. © 2015 Unciuleac and Shuman. |
| Keywords: | eukaryota; protozoa; adenylyltransferase; rna repair; deinococcus radiodurans; naegleria gruberi; mastigophora (flagellates) |
| Journal Title: | RNA |
| Volume: | 21 |
| Issue: | 5 |
| ISSN: | 1355-8382 |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Date Published: | 2015-05-01 |
| Start Page: | 824 |
| End Page: | 832 |
| Language: | English |
| DOI: | 10.1261/rna.049197.114 |
| PROVIDER: | scopus |
| PMCID: | PMC4408790 |
| PUBMED: | 25740837 |
| DOI/URL: | |
| Notes: | Export Date: 4 May 2015 -- Source: Scopus |
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