Article ; Online: Characterization of RNase J.
2023 Volume 692, Page(s) 177–215
Abstract: RNase J is involved in RNA maturation as well as degradation of RNA to the level of mononucleotides ... of the cellular metabolism in bacteria. RNase J is the first ribonuclease that was shown to have both endonuclease ... and 5'-3' exonuclease activity. RNase J enzymes can be identified by their characteristic sequence ...
Abstract | RNase J is involved in RNA maturation as well as degradation of RNA to the level of mononucleotides. This enzyme plays a vital role in maintaining intracellular RNA levels and governs different steps of the cellular metabolism in bacteria. RNase J is the first ribonuclease that was shown to have both endonuclease and 5'-3' exonuclease activity. RNase J enzymes can be identified by their characteristic sequence features and domain architecture. The quaternary structure of RNase J plays a role in regulating enzyme activity. The structure of RNase J has been characterized from several homologs. These reveal extensive overall structural similarity alongside a distinct active site topology that coordinates a metal cofactor. The metal cofactor is essential for catalytic activity. The catalytic activity of RNase J is influenced by oligomerization, the choice and stoichiometry of metal cofactors, and the 5' phosphorylation state of the RNA substrate. Here we describe the sequence and structural features of RNase J alongside phylogenetic analysis and reported functional roles in diverse organisms. We also provide a detailed purification strategy to obtain an RNase J enzyme sample with or without a metal cofactor. Different methods to identify the nature of the bound metal cofactor, the binding affinity and stoichiometry are presented. Finally, we describe enzyme assays to characterize RNase J using radioactive and fluorescence-based strategies with diverse RNA substrates. |
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MeSH term(s) | Ribonucleases/metabolism ; Phylogeny ; Endoribonucleases/metabolism ; RNA/chemistry ; Ribonuclease, Pancreatic ; Metals |
Chemical Substances | Ribonucleases (EC 3.1.-) ; Endoribonucleases (EC 3.1.-) ; RNA (63231-63-0) ; Ribonuclease, Pancreatic (EC 3.1.27.5) ; Metals |
Language | English |
Publishing date | 2023-05-01 |
Publishing country | United States |
Document type | Journal Article ; Research Support, Non-U.S. Gov't |
ISSN | 1557-7988 |
ISSN (online) | 1557-7988 |
DOI | 10.1016/bs.mie.2023.03.020 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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