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Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci

In C. elegans, efficient RNA silencing requires small RNA amplification mediated by RNA-dependent RNA polymerases (RdRPs). RRF-1, an RdRP, and other Mutator complex proteins localize to Mutator foci, which are perinuclear germline foci that associate with nuclear pores and P granules to facilitate s...

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Autores principales: Uebel, Celja J., Anderson, Dorian C., Mandarino, Lisa M., Manage, Kevin I., Aynaszyan, Stephan, Phillips, Carolyn M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072111/
https://www.ncbi.nlm.nih.gov/pubmed/30036386
http://dx.doi.org/10.1371/journal.pgen.1007542
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author Uebel, Celja J.
Anderson, Dorian C.
Mandarino, Lisa M.
Manage, Kevin I.
Aynaszyan, Stephan
Phillips, Carolyn M.
author_facet Uebel, Celja J.
Anderson, Dorian C.
Mandarino, Lisa M.
Manage, Kevin I.
Aynaszyan, Stephan
Phillips, Carolyn M.
author_sort Uebel, Celja J.
collection PubMed
description In C. elegans, efficient RNA silencing requires small RNA amplification mediated by RNA-dependent RNA polymerases (RdRPs). RRF-1, an RdRP, and other Mutator complex proteins localize to Mutator foci, which are perinuclear germline foci that associate with nuclear pores and P granules to facilitate small RNA amplification. The Mutator complex protein MUT-16 is critical for Mutator foci assembly. By analyzing small deletions of MUT-16, we identify specific regions of the protein that recruit other Mutator complex components and demonstrate that it acts as a scaffolding protein. We further determine that the C-terminal region of MUT-16, a portion of which contains predicted intrinsic disorder, is necessary and sufficient to promote Mutator foci formation. Finally, we establish that MUT-16 foci have many properties consistent with a phase-separated condensate and propose that Mutator foci form through liquid-liquid phase separation of MUT-16. P granules, which contain additional RNA silencing proteins, have previously been shown to have liquid-like properties. Thus, RNA silencing in C. elegans germ cells may rely on multiple phase-separated compartments through which sorting, processing, and silencing of mRNAs occurs.
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spelling pubmed-60721112018-08-16 Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci Uebel, Celja J. Anderson, Dorian C. Mandarino, Lisa M. Manage, Kevin I. Aynaszyan, Stephan Phillips, Carolyn M. PLoS Genet Research Article In C. elegans, efficient RNA silencing requires small RNA amplification mediated by RNA-dependent RNA polymerases (RdRPs). RRF-1, an RdRP, and other Mutator complex proteins localize to Mutator foci, which are perinuclear germline foci that associate with nuclear pores and P granules to facilitate small RNA amplification. The Mutator complex protein MUT-16 is critical for Mutator foci assembly. By analyzing small deletions of MUT-16, we identify specific regions of the protein that recruit other Mutator complex components and demonstrate that it acts as a scaffolding protein. We further determine that the C-terminal region of MUT-16, a portion of which contains predicted intrinsic disorder, is necessary and sufficient to promote Mutator foci formation. Finally, we establish that MUT-16 foci have many properties consistent with a phase-separated condensate and propose that Mutator foci form through liquid-liquid phase separation of MUT-16. P granules, which contain additional RNA silencing proteins, have previously been shown to have liquid-like properties. Thus, RNA silencing in C. elegans germ cells may rely on multiple phase-separated compartments through which sorting, processing, and silencing of mRNAs occurs. Public Library of Science 2018-07-23 /pmc/articles/PMC6072111/ /pubmed/30036386 http://dx.doi.org/10.1371/journal.pgen.1007542 Text en © 2018 Uebel et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Uebel, Celja J.
Anderson, Dorian C.
Mandarino, Lisa M.
Manage, Kevin I.
Aynaszyan, Stephan
Phillips, Carolyn M.
Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title_full Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title_fullStr Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title_full_unstemmed Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title_short Distinct regions of the intrinsically disordered protein MUT-16 mediate assembly of a small RNA amplification complex and promote phase separation of Mutator foci
title_sort distinct regions of the intrinsically disordered protein mut-16 mediate assembly of a small rna amplification complex and promote phase separation of mutator foci
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6072111/
https://www.ncbi.nlm.nih.gov/pubmed/30036386
http://dx.doi.org/10.1371/journal.pgen.1007542
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