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Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches
Previous genetic and biochemical studies from Saccharomyces cerevisiae have identified a critical ribosome-associated quality control complex (RQC) that facilitates resolution of stalled ribosomal complexes. While components of the mammalian RQC have been examined in vitro, a systematic characteriza...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5935074/ https://www.ncbi.nlm.nih.gov/pubmed/29540532 http://dx.doi.org/10.1091/mbc.E17-12-0714 |
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author | Zuzow, Nathan Ghosh, Arit Leonard, Marilyn Liao, Jeffrey Yang, Bing Bennett, Eric J. |
author_facet | Zuzow, Nathan Ghosh, Arit Leonard, Marilyn Liao, Jeffrey Yang, Bing Bennett, Eric J. |
author_sort | Zuzow, Nathan |
collection | PubMed |
description | Previous genetic and biochemical studies from Saccharomyces cerevisiae have identified a critical ribosome-associated quality control complex (RQC) that facilitates resolution of stalled ribosomal complexes. While components of the mammalian RQC have been examined in vitro, a systematic characterization of RQC protein interactions in mammalian cells has yet to be described. Here we utilize both proximity-labeling proteomic approaches, BioID and APEX, and traditional affinity-based strategies to both identify interacting proteins of mammalian RQC members and putative substrates for the RQC resident E3 ligase, Ltn1. Surprisingly, validation studies revealed that a subset of substrates are ubiquitylated by Ltn1 in a regulatory manner that does not result in subsequent substrate degradation. We demonstrate that Ltn1 catalyzes the regulatory ubiquitylation of ribosomal protein S6 kinase 1 and 2 (RPS6KA1, RPS6KA3). Further, loss of Ltn1 function results in hyperactivation of RSK1/2 signaling without impacting RSK1/2 protein turnover. These results suggest that Ltn1-mediated RSK1/2 ubiquitylation is inhibitory and establishes a new role for Ltn1 in regulating mitogen-activated kinase signaling via regulatory RSK1/2 ubiquitylation. Taken together, our results suggest that mammalian RQC interactions are difficult to observe and may be more transient than the homologous complex in S. cerevisiae and that Ltn1 has RQC-independent functions. |
format | Online Article Text |
id | pubmed-5935074 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-59350742018-07-30 Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches Zuzow, Nathan Ghosh, Arit Leonard, Marilyn Liao, Jeffrey Yang, Bing Bennett, Eric J. Mol Biol Cell Articles Previous genetic and biochemical studies from Saccharomyces cerevisiae have identified a critical ribosome-associated quality control complex (RQC) that facilitates resolution of stalled ribosomal complexes. While components of the mammalian RQC have been examined in vitro, a systematic characterization of RQC protein interactions in mammalian cells has yet to be described. Here we utilize both proximity-labeling proteomic approaches, BioID and APEX, and traditional affinity-based strategies to both identify interacting proteins of mammalian RQC members and putative substrates for the RQC resident E3 ligase, Ltn1. Surprisingly, validation studies revealed that a subset of substrates are ubiquitylated by Ltn1 in a regulatory manner that does not result in subsequent substrate degradation. We demonstrate that Ltn1 catalyzes the regulatory ubiquitylation of ribosomal protein S6 kinase 1 and 2 (RPS6KA1, RPS6KA3). Further, loss of Ltn1 function results in hyperactivation of RSK1/2 signaling without impacting RSK1/2 protein turnover. These results suggest that Ltn1-mediated RSK1/2 ubiquitylation is inhibitory and establishes a new role for Ltn1 in regulating mitogen-activated kinase signaling via regulatory RSK1/2 ubiquitylation. Taken together, our results suggest that mammalian RQC interactions are difficult to observe and may be more transient than the homologous complex in S. cerevisiae and that Ltn1 has RQC-independent functions. The American Society for Cell Biology 2018-05-15 /pmc/articles/PMC5935074/ /pubmed/29540532 http://dx.doi.org/10.1091/mbc.E17-12-0714 Text en © 2018 Zuzow et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0/ This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Articles Zuzow, Nathan Ghosh, Arit Leonard, Marilyn Liao, Jeffrey Yang, Bing Bennett, Eric J. Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title | Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title_full | Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title_fullStr | Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title_full_unstemmed | Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title_short | Mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
title_sort | mapping the mammalian ribosome quality control complex interactome using proximity labeling approaches |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5935074/ https://www.ncbi.nlm.nih.gov/pubmed/29540532 http://dx.doi.org/10.1091/mbc.E17-12-0714 |
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