Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction

Abstract Protein phosphorylation enables a rapid adjustment of cellular activities to diverse intracellular and environmental stimuli. Many phosphoproteins are targeted on more than one site, which allows the integration of multiple signals and the implementation of complex responses. However, the h...

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Autores principales:	Lisa Dengler, Mihkel Örd, Lucca M. Schwab, Mart Loog, Jennifer C. Ewald
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1923e6617fb443f2ac1c3af087fab212
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spelling	oai:doaj.org-article:1923e6617fb443f2ac1c3af087fab2122021-12-02T14:01:32ZRegulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction10.1038/s41598-020-80357-32045-2322https://doaj.org/article/1923e6617fb443f2ac1c3af087fab2122021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80357-3https://doaj.org/toc/2045-2322Abstract Protein phosphorylation enables a rapid adjustment of cellular activities to diverse intracellular and environmental stimuli. Many phosphoproteins are targeted on more than one site, which allows the integration of multiple signals and the implementation of complex responses. However, the hierarchy and interplay between multiple phospho-sites are often unknown. Here, we study multi‐site phosphorylation using the yeast trehalase Nth1 and its activator, the 14-3-3 protein Bmh1, as a model. Nth1 is known to be phosphorylated by the metabolic kinase PKA on four serine residues and by the cell cycle kinase CDK on one residue. However, how these five phospho-sites adjust Nth1 activity remains unclear. Using a novel reporter construct, we investigated the contribution of the individual sites for the regulation of the trehalase and its 14-3-3 interactor. In contrast to the constitutively phosphorylated S20 and S83, the weaker sites S21 and S60 are only phosphorylated by increased PKA activity. For binding Bmh1, S83 functions as the high‐affinity “gatekeeper” site, but successful binding of the Bmh1 dimer and thus Nth1 activation requires S60 as a secondary site. Under nutrient-poor conditions with low PKA activity, S60 is not efficiently phosphorylated and the cell cycle dependent phosphorylation of S66 by Cdk1 contributes to Nth1 activity, likely by providing an alternative Bmh1 binding site. Additionally, the PKA sites S20 and S21 modulate the dephosphorylation of Nth1 on downstream Bmh1 sites. In summary, our results expand our molecular understanding of Nth1 regulation and provide a new aspect of the interaction of 14-3-3 proteins with their targets.Lisa DenglerMihkel ÖrdLucca M. SchwabMart LoogJennifer C. EwaldNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lisa Dengler Mihkel Örd Lucca M. Schwab Mart Loog Jennifer C. Ewald Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
description	Abstract Protein phosphorylation enables a rapid adjustment of cellular activities to diverse intracellular and environmental stimuli. Many phosphoproteins are targeted on more than one site, which allows the integration of multiple signals and the implementation of complex responses. However, the hierarchy and interplay between multiple phospho-sites are often unknown. Here, we study multi‐site phosphorylation using the yeast trehalase Nth1 and its activator, the 14-3-3 protein Bmh1, as a model. Nth1 is known to be phosphorylated by the metabolic kinase PKA on four serine residues and by the cell cycle kinase CDK on one residue. However, how these five phospho-sites adjust Nth1 activity remains unclear. Using a novel reporter construct, we investigated the contribution of the individual sites for the regulation of the trehalase and its 14-3-3 interactor. In contrast to the constitutively phosphorylated S20 and S83, the weaker sites S21 and S60 are only phosphorylated by increased PKA activity. For binding Bmh1, S83 functions as the high‐affinity “gatekeeper” site, but successful binding of the Bmh1 dimer and thus Nth1 activation requires S60 as a secondary site. Under nutrient-poor conditions with low PKA activity, S60 is not efficiently phosphorylated and the cell cycle dependent phosphorylation of S66 by Cdk1 contributes to Nth1 activity, likely by providing an alternative Bmh1 binding site. Additionally, the PKA sites S20 and S21 modulate the dephosphorylation of Nth1 on downstream Bmh1 sites. In summary, our results expand our molecular understanding of Nth1 regulation and provide a new aspect of the interaction of 14-3-3 proteins with their targets.
format	article
author	Lisa Dengler Mihkel Örd Lucca M. Schwab Mart Loog Jennifer C. Ewald
author_facet	Lisa Dengler Mihkel Örd Lucca M. Schwab Mart Loog Jennifer C. Ewald
author_sort	Lisa Dengler
title	Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
title_short	Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
title_full	Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
title_fullStr	Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
title_full_unstemmed	Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
title_sort	regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1923e6617fb443f2ac1c3af087fab212
work_keys_str_mv	AT lisadengler regulationoftrehalaseactivitybymultisitephosphorylationand1433interaction AT mihkelord regulationoftrehalaseactivitybymultisitephosphorylationand1433interaction AT luccamschwab regulationoftrehalaseactivitybymultisitephosphorylationand1433interaction AT martloog regulationoftrehalaseactivitybymultisitephosphorylationand1433interaction AT jennifercewald regulationoftrehalaseactivitybymultisitephosphorylationand1433interaction
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Regulation of trehalase activity by multi-site phosphorylation and 14-3-3 interaction

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