Reprogramming the TRNA-splicing Activity of a Bacterial RNA Repair Enzyme

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2007 May 10

PMID 17488852

Citations 18

Authors

Niroshika Keppetipola

Jayakrishnan Nandakumar

Stewart Shuman

Affiliations

Soon will be listed here.

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 (CthPnkp) can catalyze both of the end-healing steps of tRNA splicing in vitro. The route of tRNA repair by CthPnkp 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 CthPnkp 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.

Citing Articles

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Ghosh S, Ejaz A, Repeta L, Shuman S Nucleic Acids Res. 2020; 49(2):1023-1032.

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Structures of bacterial polynucleotide kinase in a Michaelis complex with GTP•Mg2+ and 5'-OH oligonucleotide and a product complex with GDP•Mg2+ and 5'-PO4 oligonucleotide reveal a mechanism of general acid-base catalysis and the determinants of....

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Structural and biochemical analysis of the phosphate donor specificity of the polynucleotide kinase component of the bacterial pnkp•hen1 RNA repair system.

Das U, Wang L, Smith P, Shuman S Biochemistry. 2013; 52(27):4734-43.

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Mapping the RNA-Seq trash bin: unusual transcripts in prokaryotic transcriptome sequencing data.

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Structure and mechanism of the 2',3' phosphatase component of the bacterial Pnkp-Hen1 RNA repair system.

Wang L, Smith P, Shuman S Nucleic Acids Res. 2013; 41(11):5864-73.

PMID: 23595150 PMC: 3675462. DOI: 10.1093/nar/gkt221.

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