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Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation
MAP kinases of the ERK family are conserved from yeast to humans. Their catalytic activity is dependent on dual phosphorylation of their activation loop’s TEY motif, catalyzed by MAPK kinases (MEKs). Here we studied variants of Mpk1, a yeast orthologue of Erk, which is essential for cell wall integr...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5007096/ https://www.ncbi.nlm.nih.gov/pubmed/27413009 http://dx.doi.org/10.1091/mbc.E16-03-0167 |
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author | Goshen-Lago, Tal Goldberg-Carp, Anat Melamed, Dganit Darlyuk-Saadon, Ilona Bai, Chen Ahn, Natalie G. Admon, Arie Engelberg, David |
author_facet | Goshen-Lago, Tal Goldberg-Carp, Anat Melamed, Dganit Darlyuk-Saadon, Ilona Bai, Chen Ahn, Natalie G. Admon, Arie Engelberg, David |
author_sort | Goshen-Lago, Tal |
collection | PubMed |
description | MAP kinases of the ERK family are conserved from yeast to humans. Their catalytic activity is dependent on dual phosphorylation of their activation loop’s TEY motif, catalyzed by MAPK kinases (MEKs). Here we studied variants of Mpk1, a yeast orthologue of Erk, which is essential for cell wall integrity. Cells lacking MPK1, or the genes encoding the relevant MEKs, MKK1 and MKK2, do not proliferate under cell wall stress, imposed, for example, by caffeine. Mutants of Mpk1, Mpk1(Y268C) and Mpk1(Y268A), function independently of Mkk1 and Mkk2. We show that these variants are phosphorylated at their activation loop in mkk1∆mkk2∆ and mkk1∆mkk2∆pbs2∆ste7∆ cells, suggesting that they autophosphorylate. However, strikingly, when Y268C/A mutations were combined with the kinase-dead mutation, K54R, or mutations at the TEY motif, T190A+Y192F, the resulting proteins still allowed mkk1∆mkk2∆ cells to proliferate under caffeine stress. Mutating the equivalent residue, Tyr-280/Tyr-261, in Erk1/Erk2 significantly impaired Erk1/2’s catalytic activity. This study describes the first case in which a MAPK, Erk/Mpk1, imposes a phenotype via a mechanism that is independent of TEY phosphorylation and an unusual case in which an equivalent mutation in a highly conserved domain of yeast and mammalian Erks causes an opposite effect. |
format | Online Article Text |
id | pubmed-5007096 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-50070962016-11-16 Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation Goshen-Lago, Tal Goldberg-Carp, Anat Melamed, Dganit Darlyuk-Saadon, Ilona Bai, Chen Ahn, Natalie G. Admon, Arie Engelberg, David Mol Biol Cell Articles MAP kinases of the ERK family are conserved from yeast to humans. Their catalytic activity is dependent on dual phosphorylation of their activation loop’s TEY motif, catalyzed by MAPK kinases (MEKs). Here we studied variants of Mpk1, a yeast orthologue of Erk, which is essential for cell wall integrity. Cells lacking MPK1, or the genes encoding the relevant MEKs, MKK1 and MKK2, do not proliferate under cell wall stress, imposed, for example, by caffeine. Mutants of Mpk1, Mpk1(Y268C) and Mpk1(Y268A), function independently of Mkk1 and Mkk2. We show that these variants are phosphorylated at their activation loop in mkk1∆mkk2∆ and mkk1∆mkk2∆pbs2∆ste7∆ cells, suggesting that they autophosphorylate. However, strikingly, when Y268C/A mutations were combined with the kinase-dead mutation, K54R, or mutations at the TEY motif, T190A+Y192F, the resulting proteins still allowed mkk1∆mkk2∆ cells to proliferate under caffeine stress. Mutating the equivalent residue, Tyr-280/Tyr-261, in Erk1/Erk2 significantly impaired Erk1/2’s catalytic activity. This study describes the first case in which a MAPK, Erk/Mpk1, imposes a phenotype via a mechanism that is independent of TEY phosphorylation and an unusual case in which an equivalent mutation in a highly conserved domain of yeast and mammalian Erks causes an opposite effect. The American Society for Cell Biology 2016-09-01 /pmc/articles/PMC5007096/ /pubmed/27413009 http://dx.doi.org/10.1091/mbc.E16-03-0167 Text en © 2016 Goshen-Lago, Goldberg-Carp, et al. 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 (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Goshen-Lago, Tal Goldberg-Carp, Anat Melamed, Dganit Darlyuk-Saadon, Ilona Bai, Chen Ahn, Natalie G. Admon, Arie Engelberg, David Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title | Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title_full | Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title_fullStr | Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title_full_unstemmed | Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title_short | Variants of the yeast MAPK Mpk1 are fully functional independently of activation loop phosphorylation |
title_sort | variants of the yeast mapk mpk1 are fully functional independently of activation loop phosphorylation |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5007096/ https://www.ncbi.nlm.nih.gov/pubmed/27413009 http://dx.doi.org/10.1091/mbc.E16-03-0167 |
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