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Novel perspectives of target-binding by the evolutionarily conserved PP4 phosphatase

Karman, Zoltan and Rethi-Nagy, Zsuzsanna and Abraham, Edit and Fabri-Ordogh, Lilla and Csonka, Akos and Vilmos, Peter and Debski, Janusz and Dadlez, Michal and Glover, David M. and Lipinszki, Zoltan (2020) Novel perspectives of target-binding by the evolutionarily conserved PP4 phosphatase. Open Biology, 10 (12). Art. No. 200343. ISSN 2046-2441. PMCID PMC7776573. https://resolver.caltech.edu/CaltechAUTHORS:20201224-085808015

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Abstract

Protein phosphatase 4 (PP4) is an evolutionarily conserved and essential Ser/Thr phosphatase that regulates cell division, development and DNA repair in eukaryotes. The major form of PP4, present from yeast to human, is the PP4c-R2-R3 heterotrimeric complex. The R3 subunit is responsible for substrate-recognition via its EVH1 domain. In typical EVH1 domains, conserved phenylalanine, tyrosine and tryptophan residues form the specific recognition site for their target's proline-rich sequences. Here, we identify novel binding partners of the EVH1 domain of the Drosophila R3 subunit, Falafel, and demonstrate that instead of binding to proline-rich sequences this EVH1 variant specifically recognizes atypical ligands, namely the FxxP and MxPP short linear consensus motifs. This interaction is dependent on an exclusively conserved leucine that replaces the phenylalanine invariant of all canonical EVH1 domains. We propose that the EVH1 domain of PP4 represents a new class of the EVH1 family that can accommodate low proline content sequences, such as the FxxP motif. Finally, our data implicate the conserved Smk-1 domain of Falafel in target-binding. These findings greatly enhance our understanding of the substrate-recognition mechanisms and function of PP4.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1098/rsob.200343DOIArticle
https://doi.org/10.6084/m9.figshare.c.5230606Related ItemSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7776573PubMed CentralArticle
ORCID:
AuthorORCID
Fabri-Ordogh, Lilla0000-0003-0405-4017
Csonka, Akos0000-0002-9628-6095
Vilmos, Peter0000-0001-5692-8818
Debski, Janusz0000-0002-0171-7797
Dadlez, Michal0000-0001-8811-5176
Glover, David M.0000-0003-0956-0103
Lipinszki, Zoltan0000-0002-2067-0832
Additional Information:© 2020 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited. Manuscript received 19/10/2020. Manuscript accepted 27/11/2020. Published online 23/12/2020. The authors thank Andor Udvardy (BRC, Hungary) and Matthew S. Savoian (Massey University, NZ) for their help and critical reading of the manuscript and Dora Bokor for proofreading the manuscript. We are also grateful to the Drosophila Genomics Resource Center (supported by NIH grant no. 2P40OD010949) for the Gold collection cDNA clones and Drosophila Gateway plasmids as well as FlyBase, the Drosophila genetic and genomic database (supported by NIH grant no. U41 HG000739 and British Medical Research Council grant no. MR/N030117/1). This work was supported by the Ministry of Human Capacities of Hungary (UNKP-18-2 and NTP-NFTÖ-19-B) and Aron Marton College to Z.R.-N., Government of Hungary (PhD scholarship) to Z.K. and Z.R.-N., Ministry for National Economy of Hungary (GINOP-2.3.2-15-2016-00001 and GINOP-2.3.2-15-2016-00032) to Z.L. and P.V., The National Research, Development and Innovation Office (OTKA-PD115404) and Hungarian Academy of Sciences (Bolyai Fellowship and Lendület grant no. (LP2017-7/2017)) to Z.L. D.M.G. is grateful for grants from Wellcome Trust (RG84496) and NIH (R01NS113930). Authors' contributions. Z.K., Z.R.-N. and Z.L. conceived the study, designed and performed experiments and wrote the manuscript. D.M.G and Z.L. supervised the project. Z.K. performed affinity purifications for mass spectrometry and with Z.R.-N. designed and carried out the binding studies. E.A. purified recombinant proteins, L.F.-O. and A.C. generated DNA constructs and ran predictions, P.V. carried out the fly work and J.D. and M.D. performed mass spectrometry. Data accessibility. The raw data are available from the authors. The authors declare that they have no competing interests.
Funders:
Funding AgencyGrant Number
NIH2P40OD010949
NIHU41 HG000739
Medical Research Council (UK)MR/N030117/1
Ministry of Human Capacities (Hungary)UNKP-18-2
Ministry of Human Capacities (Hungary)NTP-NFTÖ-19-B
Aron Marton CollegeUNSPECIFIED
National Research, Development and Innovation Office Hungary (NKFI)GINOP-2.3.2-15-2016-00001
National Research, Development and Innovation Office Hungary (NKFI)GINOP-2.3.2-15-2016-00032
Hungarian Scientific Research Fund (OTKA)OTKA-PD115404
Hungarian Academy of SciencesLP2017-7/2017
Wellcome TrustRG84496
NIHR01NS113930
Subject Keywords:binding motif; EVH1; SLiM; Drosophila; Smk-1; PP4
Issue or Number:12
PubMed Central ID:PMC7776573
Record Number:CaltechAUTHORS:20201224-085808015
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201224-085808015
Official Citation:Karman Zoltan, Rethi-Nagy Zsuzsanna, Abraham Edit, Fabri-Ordogh Lilla, Csonka Akos, Vilmos Peter, Debski Janusz, Dadlez Michal, Glover David M. and Lipinszki Zoltan 2020 Novel perspectives of target-binding by the evolutionarily conserved PP4 phosphatase Open Biol.10200343 http://doi.org/10.1098/rsob.200343
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107280
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:04 Jan 2021 16:19
Last Modified:11 Jan 2021 17:44

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