Cargando…
Tagging the proteasome active site β5 causes tag specific phenotypes in yeast
The efficient and timely degradation of proteins is crucial for many cellular processes and to maintain general proteostasis. The proteasome, a complex multisubunit protease, plays a critical role in protein degradation. Therefore, it is important to understand the assembly, regulation, and localiza...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582879/ https://www.ncbi.nlm.nih.gov/pubmed/33093623 http://dx.doi.org/10.1038/s41598-020-75126-1 |
_version_ | 1783599292422815744 |
---|---|
author | Waite, Kenrick A. Burris, Alicia Roelofs, Jeroen |
author_facet | Waite, Kenrick A. Burris, Alicia Roelofs, Jeroen |
author_sort | Waite, Kenrick A. |
collection | PubMed |
description | The efficient and timely degradation of proteins is crucial for many cellular processes and to maintain general proteostasis. The proteasome, a complex multisubunit protease, plays a critical role in protein degradation. Therefore, it is important to understand the assembly, regulation, and localization of proteasome complexes in the cell under different conditions. Fluorescent tags are often utilized to study proteasomes. A GFP-tag on the β5 subunit, one of the core particle (CP) subunits with catalytic activity, has been shown to be incorporated into proteasomes and commonly used by the field. We report here that a tag on this subunit results in aberrant phenotypes that are not observed when several other CP subunits are tagged. These phenotypes appear in combination with other proteasome mutations and include poor growth, and, more significantly, altered 26S proteasome localization. In strains defective for autophagy, β5-GFP tagged proteasomes, unlike other CP tags, localize to granules upon nitrogen starvation. These granules are reflective of previously described proteasome storage granules but display unique properties. This suggests proteasomes with a β5-GFP tag are specifically recognized and sequestered depending on physiological conditions. In all, our data indicate the intricacy of tagging proteasomes, and possibly, large complexes in general. |
format | Online Article Text |
id | pubmed-7582879 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-75828792020-10-23 Tagging the proteasome active site β5 causes tag specific phenotypes in yeast Waite, Kenrick A. Burris, Alicia Roelofs, Jeroen Sci Rep Article The efficient and timely degradation of proteins is crucial for many cellular processes and to maintain general proteostasis. The proteasome, a complex multisubunit protease, plays a critical role in protein degradation. Therefore, it is important to understand the assembly, regulation, and localization of proteasome complexes in the cell under different conditions. Fluorescent tags are often utilized to study proteasomes. A GFP-tag on the β5 subunit, one of the core particle (CP) subunits with catalytic activity, has been shown to be incorporated into proteasomes and commonly used by the field. We report here that a tag on this subunit results in aberrant phenotypes that are not observed when several other CP subunits are tagged. These phenotypes appear in combination with other proteasome mutations and include poor growth, and, more significantly, altered 26S proteasome localization. In strains defective for autophagy, β5-GFP tagged proteasomes, unlike other CP tags, localize to granules upon nitrogen starvation. These granules are reflective of previously described proteasome storage granules but display unique properties. This suggests proteasomes with a β5-GFP tag are specifically recognized and sequestered depending on physiological conditions. In all, our data indicate the intricacy of tagging proteasomes, and possibly, large complexes in general. Nature Publishing Group UK 2020-10-22 /pmc/articles/PMC7582879/ /pubmed/33093623 http://dx.doi.org/10.1038/s41598-020-75126-1 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Waite, Kenrick A. Burris, Alicia Roelofs, Jeroen Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title | Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title_full | Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title_fullStr | Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title_full_unstemmed | Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title_short | Tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
title_sort | tagging the proteasome active site β5 causes tag specific phenotypes in yeast |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582879/ https://www.ncbi.nlm.nih.gov/pubmed/33093623 http://dx.doi.org/10.1038/s41598-020-75126-1 |
work_keys_str_mv | AT waitekenricka taggingtheproteasomeactivesiteb5causestagspecificphenotypesinyeast AT burrisalicia taggingtheproteasomeactivesiteb5causestagspecificphenotypesinyeast AT roelofsjeroen taggingtheproteasomeactivesiteb5causestagspecificphenotypesinyeast |