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Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes

Mitochondrial tRNAs are transcribed as long polycistronic transcripts of precursor tRNAs and undergo posttranscriptional modifications such as endonucleolytic processing and methylation required for their correct structure and function. Among them, 5′-end processing and purine 9 N1-methylation of mi...

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Autores principales: Oerum, Stephanie, Roovers, Martine, Rambo, Robert P., Kopec, Jola, Bailey, Henry J., Fitzpatrick, Fiona, Newman, Joseph A., Newman, William G., Amberger, Albert, Zschocke, Johannes, Droogmans, Louis, Oppermann, Udo, Yue, Wyatt W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6102140/
https://www.ncbi.nlm.nih.gov/pubmed/29880640
http://dx.doi.org/10.1074/jbc.RA117.001286
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author Oerum, Stephanie
Roovers, Martine
Rambo, Robert P.
Kopec, Jola
Bailey, Henry J.
Fitzpatrick, Fiona
Newman, Joseph A.
Newman, William G.
Amberger, Albert
Zschocke, Johannes
Droogmans, Louis
Oppermann, Udo
Yue, Wyatt W.
author_facet Oerum, Stephanie
Roovers, Martine
Rambo, Robert P.
Kopec, Jola
Bailey, Henry J.
Fitzpatrick, Fiona
Newman, Joseph A.
Newman, William G.
Amberger, Albert
Zschocke, Johannes
Droogmans, Louis
Oppermann, Udo
Yue, Wyatt W.
author_sort Oerum, Stephanie
collection PubMed
description Mitochondrial tRNAs are transcribed as long polycistronic transcripts of precursor tRNAs and undergo posttranscriptional modifications such as endonucleolytic processing and methylation required for their correct structure and function. Among them, 5′-end processing and purine 9 N1-methylation of mitochondrial tRNA are catalyzed by two proteinaceous complexes with overlapping subunit composition. The Mg(2+)-dependent RNase P complex for 5′-end cleavage comprises the methyltransferase domain–containing protein tRNA methyltransferase 10C, mitochondrial RNase P subunit (TRMT10C/MRPP1), short-chain oxidoreductase hydroxysteroid 17β-dehydrogenase 10 (HSD17B10/MRPP2), and metallonuclease KIAA0391/MRPP3. An MRPP1–MRPP2 subcomplex also catalyzes the formation of 1-methyladenosine/1-methylguanosine at position 9 using S-adenosyl-l-methionine as methyl donor. However, a lack of structural information has precluded insights into how these complexes methylate and process mitochondrial tRNA. Here, we used a combination of X-ray crystallography, interaction and activity assays, and small angle X-ray scattering (SAXS) to gain structural insight into the two tRNA modification complexes and their components. The MRPP1 N terminus is involved in tRNA binding and monomer–monomer self-interaction, whereas the C-terminal SPOUT fold contains key residues for S-adenosyl-l-methionine binding and N1-methylation. The entirety of MRPP1 interacts with MRPP2 to form the N1-methylation complex, whereas the MRPP1–MRPP2–MRPP3 RNase P complex only assembles in the presence of precursor tRNA. This study proposes low-resolution models of the MRPP1–MRPP2 and MRPP1–MRPP2–MRPP3 complexes that suggest the overall architecture, stoichiometry, and orientation of subunits and tRNA substrates.
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spelling pubmed-61021402018-08-21 Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes Oerum, Stephanie Roovers, Martine Rambo, Robert P. Kopec, Jola Bailey, Henry J. Fitzpatrick, Fiona Newman, Joseph A. Newman, William G. Amberger, Albert Zschocke, Johannes Droogmans, Louis Oppermann, Udo Yue, Wyatt W. J Biol Chem Protein Structure and Folding Mitochondrial tRNAs are transcribed as long polycistronic transcripts of precursor tRNAs and undergo posttranscriptional modifications such as endonucleolytic processing and methylation required for their correct structure and function. Among them, 5′-end processing and purine 9 N1-methylation of mitochondrial tRNA are catalyzed by two proteinaceous complexes with overlapping subunit composition. The Mg(2+)-dependent RNase P complex for 5′-end cleavage comprises the methyltransferase domain–containing protein tRNA methyltransferase 10C, mitochondrial RNase P subunit (TRMT10C/MRPP1), short-chain oxidoreductase hydroxysteroid 17β-dehydrogenase 10 (HSD17B10/MRPP2), and metallonuclease KIAA0391/MRPP3. An MRPP1–MRPP2 subcomplex also catalyzes the formation of 1-methyladenosine/1-methylguanosine at position 9 using S-adenosyl-l-methionine as methyl donor. However, a lack of structural information has precluded insights into how these complexes methylate and process mitochondrial tRNA. Here, we used a combination of X-ray crystallography, interaction and activity assays, and small angle X-ray scattering (SAXS) to gain structural insight into the two tRNA modification complexes and their components. The MRPP1 N terminus is involved in tRNA binding and monomer–monomer self-interaction, whereas the C-terminal SPOUT fold contains key residues for S-adenosyl-l-methionine binding and N1-methylation. The entirety of MRPP1 interacts with MRPP2 to form the N1-methylation complex, whereas the MRPP1–MRPP2–MRPP3 RNase P complex only assembles in the presence of precursor tRNA. This study proposes low-resolution models of the MRPP1–MRPP2 and MRPP1–MRPP2–MRPP3 complexes that suggest the overall architecture, stoichiometry, and orientation of subunits and tRNA substrates. American Society for Biochemistry and Molecular Biology 2018-08-17 2018-06-07 /pmc/articles/PMC6102140/ /pubmed/29880640 http://dx.doi.org/10.1074/jbc.RA117.001286 Text en © 2018 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Protein Structure and Folding
Oerum, Stephanie
Roovers, Martine
Rambo, Robert P.
Kopec, Jola
Bailey, Henry J.
Fitzpatrick, Fiona
Newman, Joseph A.
Newman, William G.
Amberger, Albert
Zschocke, Johannes
Droogmans, Louis
Oppermann, Udo
Yue, Wyatt W.
Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title_full Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title_fullStr Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title_full_unstemmed Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title_short Structural insight into the human mitochondrial tRNA purine N1-methyltransferase and ribonuclease P complexes
title_sort structural insight into the human mitochondrial trna purine n1-methyltransferase and ribonuclease p complexes
topic Protein Structure and Folding
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6102140/
https://www.ncbi.nlm.nih.gov/pubmed/29880640
http://dx.doi.org/10.1074/jbc.RA117.001286
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