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Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex

Grc3 is an essential well-conserved eukaryotic polynucleotide kinase (PNK) that cooperates with the endoribonuclease Las1 to process the preribosomal RNA (rRNA). Aside from being dependent upon Las1 for coordinated kinase and nuclease function, little is known about Grc3 substrate specificity and th...

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Detalles Bibliográficos
Autores principales: Pillon, Monica C., Sobhany, Mack, Stanley, Robin E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5900568/
https://www.ncbi.nlm.nih.gov/pubmed/29440475
http://dx.doi.org/10.1261/rna.065037.117
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author Pillon, Monica C.
Sobhany, Mack
Stanley, Robin E.
author_facet Pillon, Monica C.
Sobhany, Mack
Stanley, Robin E.
author_sort Pillon, Monica C.
collection PubMed
description Grc3 is an essential well-conserved eukaryotic polynucleotide kinase (PNK) that cooperates with the endoribonuclease Las1 to process the preribosomal RNA (rRNA). Aside from being dependent upon Las1 for coordinated kinase and nuclease function, little is known about Grc3 substrate specificity and the molecular mechanisms governing kinase activity. Here we characterize the kinase activity of Grc3 and identify key similarities and differences between Grc3 and other polynucleotide kinase family members. In contrast to other PNK family members, Grc3 has distinct substrate preference for RNA substrates in vitro. By disrupting conserved residues found at the Grc3 kinase active site, we identified specific residues required to support Grc3-directed Las1-mediated pre-rRNA cleavage in vitro and in vivo. The crosstalk between Grc3 and Las1 ensures the direct coupling of cleavage and phosphorylation during pre-rRNA processing. Taken together, our studies provide key insight into the polynucleotide kinase activity of the essential enzyme Grc3 and its molecular crosstalk with the endoribonuclease Las1.
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spelling pubmed-59005682019-05-01 Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex Pillon, Monica C. Sobhany, Mack Stanley, Robin E. RNA Article Grc3 is an essential well-conserved eukaryotic polynucleotide kinase (PNK) that cooperates with the endoribonuclease Las1 to process the preribosomal RNA (rRNA). Aside from being dependent upon Las1 for coordinated kinase and nuclease function, little is known about Grc3 substrate specificity and the molecular mechanisms governing kinase activity. Here we characterize the kinase activity of Grc3 and identify key similarities and differences between Grc3 and other polynucleotide kinase family members. In contrast to other PNK family members, Grc3 has distinct substrate preference for RNA substrates in vitro. By disrupting conserved residues found at the Grc3 kinase active site, we identified specific residues required to support Grc3-directed Las1-mediated pre-rRNA cleavage in vitro and in vivo. The crosstalk between Grc3 and Las1 ensures the direct coupling of cleavage and phosphorylation during pre-rRNA processing. Taken together, our studies provide key insight into the polynucleotide kinase activity of the essential enzyme Grc3 and its molecular crosstalk with the endoribonuclease Las1. Cold Spring Harbor Laboratory Press 2018-05 /pmc/articles/PMC5900568/ /pubmed/29440475 http://dx.doi.org/10.1261/rna.065037.117 Text en Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This is a work of the US Government.
spellingShingle Article
Pillon, Monica C.
Sobhany, Mack
Stanley, Robin E.
Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title_full Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title_fullStr Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title_full_unstemmed Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title_short Characterization of the molecular crosstalk within the essential Grc3/Las1 pre-rRNA processing complex
title_sort characterization of the molecular crosstalk within the essential grc3/las1 pre-rrna processing complex
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5900568/
https://www.ncbi.nlm.nih.gov/pubmed/29440475
http://dx.doi.org/10.1261/rna.065037.117
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