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DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma

The two p53 homologues p63 and p73 regulate transcriptional programs in epithelial tissues and several cell types in these tissues express both proteins. All members of the p53 family form tetramers in their active state through a dedicated oligomerization domain that structurally assembles as a dim...

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Autores principales: Strubel, Alexander, Münick, Philipp, Hartmann, Oliver, Chaikuad, Apirat, Dreier, Birgit, Schaefer, Jonas V., Gebel, Jakob, Osterburg, Christian, Tuppi, Marcel, Schäfer, Birgit, Buck, Viktoria, Rosenfeldt, Mathias, Knapp, Stefan, Plückthun, Andreas, Diefenbacher, Markus E., Dötsch, Volker
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10570377/
https://www.ncbi.nlm.nih.gov/pubmed/37828008
http://dx.doi.org/10.1038/s41419-023-06213-0
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author Strubel, Alexander
Münick, Philipp
Hartmann, Oliver
Chaikuad, Apirat
Dreier, Birgit
Schaefer, Jonas V.
Gebel, Jakob
Osterburg, Christian
Tuppi, Marcel
Schäfer, Birgit
Buck, Viktoria
Rosenfeldt, Mathias
Knapp, Stefan
Plückthun, Andreas
Diefenbacher, Markus E.
Dötsch, Volker
author_facet Strubel, Alexander
Münick, Philipp
Hartmann, Oliver
Chaikuad, Apirat
Dreier, Birgit
Schaefer, Jonas V.
Gebel, Jakob
Osterburg, Christian
Tuppi, Marcel
Schäfer, Birgit
Buck, Viktoria
Rosenfeldt, Mathias
Knapp, Stefan
Plückthun, Andreas
Diefenbacher, Markus E.
Dötsch, Volker
author_sort Strubel, Alexander
collection PubMed
description The two p53 homologues p63 and p73 regulate transcriptional programs in epithelial tissues and several cell types in these tissues express both proteins. All members of the p53 family form tetramers in their active state through a dedicated oligomerization domain that structurally assembles as a dimer of dimers. The oligomerization domain of p63 and p73 share a high sequence identity, but the p53 oligomerization domain is more divergent and it lacks a functionally important C-terminal helix present in the other two family members. Based on these structural differences, p53 does not hetero-oligomerize with p63 or p73. In contrast, p63 and p73 form hetero-oligomers of all possible stoichiometries, with the hetero-tetramer built from a p63 dimer and a p73 dimer being thermodynamically more stable than the two homo-tetramers. This predicts that in cells expressing both proteins a p63(2)/p73(2) hetero-tetramer is formed. So far, the tools to investigate the biological function of this hetero-tetramer have been missing. Here we report the generation and characterization of Designed Ankyrin Repeat Proteins (DARPins) that bind with high affinity and selectivity to the p63(2)/p73(2) hetero-tetramer. Using these DARPins we were able to confirm experimentally the existence of this hetero-tetramer in epithelial mouse and human tissues and show that its level increases in squamous cell carcinoma.
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spelling pubmed-105703772023-10-14 DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma Strubel, Alexander Münick, Philipp Hartmann, Oliver Chaikuad, Apirat Dreier, Birgit Schaefer, Jonas V. Gebel, Jakob Osterburg, Christian Tuppi, Marcel Schäfer, Birgit Buck, Viktoria Rosenfeldt, Mathias Knapp, Stefan Plückthun, Andreas Diefenbacher, Markus E. Dötsch, Volker Cell Death Dis Article The two p53 homologues p63 and p73 regulate transcriptional programs in epithelial tissues and several cell types in these tissues express both proteins. All members of the p53 family form tetramers in their active state through a dedicated oligomerization domain that structurally assembles as a dimer of dimers. The oligomerization domain of p63 and p73 share a high sequence identity, but the p53 oligomerization domain is more divergent and it lacks a functionally important C-terminal helix present in the other two family members. Based on these structural differences, p53 does not hetero-oligomerize with p63 or p73. In contrast, p63 and p73 form hetero-oligomers of all possible stoichiometries, with the hetero-tetramer built from a p63 dimer and a p73 dimer being thermodynamically more stable than the two homo-tetramers. This predicts that in cells expressing both proteins a p63(2)/p73(2) hetero-tetramer is formed. So far, the tools to investigate the biological function of this hetero-tetramer have been missing. Here we report the generation and characterization of Designed Ankyrin Repeat Proteins (DARPins) that bind with high affinity and selectivity to the p63(2)/p73(2) hetero-tetramer. Using these DARPins we were able to confirm experimentally the existence of this hetero-tetramer in epithelial mouse and human tissues and show that its level increases in squamous cell carcinoma. Nature Publishing Group UK 2023-10-12 /pmc/articles/PMC10570377/ /pubmed/37828008 http://dx.doi.org/10.1038/s41419-023-06213-0 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Strubel, Alexander
Münick, Philipp
Hartmann, Oliver
Chaikuad, Apirat
Dreier, Birgit
Schaefer, Jonas V.
Gebel, Jakob
Osterburg, Christian
Tuppi, Marcel
Schäfer, Birgit
Buck, Viktoria
Rosenfeldt, Mathias
Knapp, Stefan
Plückthun, Andreas
Diefenbacher, Markus E.
Dötsch, Volker
DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title_full DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title_fullStr DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title_full_unstemmed DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title_short DARPins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
title_sort darpins detect the formation of hetero-tetramers of p63 and p73 in epithelial tissues and in squamous cell carcinoma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10570377/
https://www.ncbi.nlm.nih.gov/pubmed/37828008
http://dx.doi.org/10.1038/s41419-023-06213-0
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