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Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro

Intrinsically disordered proteins (IDPs) are often multifunctional and frequently posttranslationally modified. Deleted in split hand/split foot 1 (Dss1—Sem1 in budding yeast) is a highly multifunctional IDP associated with a range of protein complexes. However, it remains unknown if the different f...

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Autores principales: Jacobsen, Nina L., Bloch, Magnus, Millard, Peter S., Ruidiaz, Sarah F., Elsborg, Jonas D., Boomsma, Wouter, Hendus‐Altenburger, Ruth, Hartmann‐Petersen, Rasmus, Kragelund, Birthe B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley & Sons, Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10443397/
https://www.ncbi.nlm.nih.gov/pubmed/37463013
http://dx.doi.org/10.1002/pro.4733
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author Jacobsen, Nina L.
Bloch, Magnus
Millard, Peter S.
Ruidiaz, Sarah F.
Elsborg, Jonas D.
Boomsma, Wouter
Hendus‐Altenburger, Ruth
Hartmann‐Petersen, Rasmus
Kragelund, Birthe B.
author_facet Jacobsen, Nina L.
Bloch, Magnus
Millard, Peter S.
Ruidiaz, Sarah F.
Elsborg, Jonas D.
Boomsma, Wouter
Hendus‐Altenburger, Ruth
Hartmann‐Petersen, Rasmus
Kragelund, Birthe B.
author_sort Jacobsen, Nina L.
collection PubMed
description Intrinsically disordered proteins (IDPs) are often multifunctional and frequently posttranslationally modified. Deleted in split hand/split foot 1 (Dss1—Sem1 in budding yeast) is a highly multifunctional IDP associated with a range of protein complexes. However, it remains unknown if the different functions relate to different modified states. In this work, we show that Schizosaccharomyces pombe Dss1 is a substrate for casein kinase 2 in vitro, and we identify three phosphorylated threonines in its linker region separating two known disordered ubiquitin‐binding motifs. Phosphorylations of the threonines had no effect on ubiquitin‐binding but caused a slight destabilization of the C‐terminal α‐helix and mediated a direct interaction with the forkhead‐associated (FHA) domain of the RING‐FHA E3‐ubiquitin ligase defective in mitosis 1 (Dma1). The phosphorylation sites are not conserved and are absent in human Dss1. Sequence analyses revealed that the Txx(E/D) motif, which is important for phosphorylation and Dma1 binding, is not linked to certain branches of the evolutionary tree. Instead, we find that the motif appears randomly, supporting the mechanism of ex nihilo evolution of novel motifs. In support of this, other threonine‐based motifs, although frequent, are nonconserved in the linker, pointing to additional functions connected to this region. We suggest that Dss1 acts as an adaptor protein that docks to Dma1 via the phosphorylated FHA‐binding motifs, while the C‐terminal α‐helix is free to bind mitotic septins, thereby stabilizing the complex. The presence of Txx(D/E) motifs in the disordered regions of certain septin subunits may be of further relevance to the formation and stabilization of these complexes.
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spelling pubmed-104433972023-09-01 Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro Jacobsen, Nina L. Bloch, Magnus Millard, Peter S. Ruidiaz, Sarah F. Elsborg, Jonas D. Boomsma, Wouter Hendus‐Altenburger, Ruth Hartmann‐Petersen, Rasmus Kragelund, Birthe B. Protein Sci Research Articles Intrinsically disordered proteins (IDPs) are often multifunctional and frequently posttranslationally modified. Deleted in split hand/split foot 1 (Dss1—Sem1 in budding yeast) is a highly multifunctional IDP associated with a range of protein complexes. However, it remains unknown if the different functions relate to different modified states. In this work, we show that Schizosaccharomyces pombe Dss1 is a substrate for casein kinase 2 in vitro, and we identify three phosphorylated threonines in its linker region separating two known disordered ubiquitin‐binding motifs. Phosphorylations of the threonines had no effect on ubiquitin‐binding but caused a slight destabilization of the C‐terminal α‐helix and mediated a direct interaction with the forkhead‐associated (FHA) domain of the RING‐FHA E3‐ubiquitin ligase defective in mitosis 1 (Dma1). The phosphorylation sites are not conserved and are absent in human Dss1. Sequence analyses revealed that the Txx(E/D) motif, which is important for phosphorylation and Dma1 binding, is not linked to certain branches of the evolutionary tree. Instead, we find that the motif appears randomly, supporting the mechanism of ex nihilo evolution of novel motifs. In support of this, other threonine‐based motifs, although frequent, are nonconserved in the linker, pointing to additional functions connected to this region. We suggest that Dss1 acts as an adaptor protein that docks to Dma1 via the phosphorylated FHA‐binding motifs, while the C‐terminal α‐helix is free to bind mitotic septins, thereby stabilizing the complex. The presence of Txx(D/E) motifs in the disordered regions of certain septin subunits may be of further relevance to the formation and stabilization of these complexes. John Wiley & Sons, Inc. 2023-09-01 /pmc/articles/PMC10443397/ /pubmed/37463013 http://dx.doi.org/10.1002/pro.4733 Text en © 2023 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Jacobsen, Nina L.
Bloch, Magnus
Millard, Peter S.
Ruidiaz, Sarah F.
Elsborg, Jonas D.
Boomsma, Wouter
Hendus‐Altenburger, Ruth
Hartmann‐Petersen, Rasmus
Kragelund, Birthe B.
Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title_full Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title_fullStr Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title_full_unstemmed Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title_short Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin‐ligase Dma1 in vitro
title_sort phosphorylation of schizosaccharomyces pombe dss1 mediates direct binding to the ubiquitin‐ligase dma1 in vitro
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10443397/
https://www.ncbi.nlm.nih.gov/pubmed/37463013
http://dx.doi.org/10.1002/pro.4733
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