Reprogramming the tRNA-splicing activity of a bacterial RNA repair enzyme Journal Article
| Authors: | Keppetipola, N.; Nandakumar, J.; Shuman, S. |
| Article Title: | Reprogramming the tRNA-splicing activity of a bacterial RNA repair enzyme |
| Abstract: | Programmed RNA breakage is an emerging theme underlying cellular responses to stress, virus infection and defense against foreign species. In many cases, site-specific cleavage of the target RNA generates 2′ 3′ cyclic phosphate and 5′-OH ends. For the damage to be repaired, both broken ends must be healed before they can be sealed by a ligase. Healing entails hydrolysis of the 2′,3′ cyclic phosphate to form a 3′-OH and phosphorylation of the 5′-OH to form a 5′-PO4. Here, we demonstrate that a polynucleotide kinase-phosphatase enzyme from Clostridium thermocellum (Cth Pnkp) can catalyze both of the end-healing steps of tRNA splicing in vitro. The route of tRNA repair by Cth Pnkp can be reprogrammed by a mutation in the 3′ end-healing domain (H189D) that yields a 2′-PO4 product instead of a 2′-OH. Whereas tRNA ends healed by wild-type Cth Pnkp are readily sealed by T4 RNA ligase 1, the H189D enzyme generates ends that are spliced by yeast tRNA ligase. Our findings suggest that RNA repair enzymes can evolve their specificities to suit a particular pathway. © 2007 The Author(s). |
| Keywords: | signal transduction; controlled study; unclassified drug; gene mutation; genetics; nonhuman; protein domain; metabolism; amino acid substitution; phosphatase; in vitro study; enzymology; wild type; bacteria (microorganisms); bacterial protein; chemistry; bacterial proteins; 3' untranslated region; catalysis; transfer rna; rna, transfer; enzyme specificity; phosphoric monoester hydrolases; aspartic acid; histidine; rna splicing; bacterial genetics; rna repair; bacterial enzyme; clostridium thermocellum; bacterial rna; rna ligase; polynucleotide kinase phosphatase |
| Journal Title: | Nucleic Acids Research |
| Volume: | 35 |
| Issue: | 11 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2007-01-01 |
| Start Page: | 3624 |
| End Page: | 3630 |
| Language: | English |
| DOI: | 10.1093/nar/gkm110 |
| PUBMED: | 17488852 |
| PROVIDER: | scopus |
| PMCID: | PMC1920235 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 13" - "Export Date: 17 November 2011" - "CODEN: NARHA" - "Source: Scopus" |
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546Shuman
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