14-3-3 Binding Motif Phosphorylation Disrupts Hdac4-organized Condensates to Stimulate Cardiac Reprogramming

Overview

Journal Cell Rep

Publisher Cell Press

Specialties Biology
Cell Biology
Molecular Biology

Date 2024 Apr 5

PMID 38578832

Authors

Liu Liu

Ienglam Lei

Shuo Tian

Wenbin Gao

Yijing Guo

Zhaokai Li

Ziad Sabry

Paul Tang

Y Eugene Chen

Zhong Wang

Affiliations

Soon will be listed here.

Abstract

Cell fate conversion is associated with extensive post-translational modifications (PTMs) and architectural changes of sub-organelles, yet how these events are interconnected remains unknown. We report here the identification of a phosphorylation code in 14-3-3 binding motifs (PC14-3-3) that greatly stimulates induced cardiomyocyte (iCM) formation from fibroblasts. PC14-3-3 is identified in pivotal functional proteins for iCM reprogramming, including transcription factors and chromatin modifiers. Akt1 kinase and protein phosphatase 2A are the key writer and key eraser of the PC14-3-3 code, respectively. PC14-3-3 activation induces iCM formation with the presence of only Tbx5. In contrast, PC14-3-3 inhibition by mutagenesis or inhibitor-mediated code removal abolishes reprogramming. We discover that key PC14-3-3-embedded factors, such as histone deacetylase 4 (Hdac4), Mef2c, and Foxo1, form Hdac4-organized inhibitory nuclear condensates. PC14-3-3 activation disrupts Hdac4 condensates to promote cardiac gene expression. Our study suggests that sub-organelle dynamics regulated by a PTM code could be a general mechanism for stimulating cell reprogramming.

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References

Oates M, Romero P, Ishida T, Ghalwash M, Mizianty M, Xue B . D²P²: database of disordered protein predictions. Nucleic Acids Res. 2012; 41(Database issue):D508-16. PMC: 3531159. DOI: 10.1093/nar/gks1226. View

Zhou H, Dickson M, Kim M, Bassel-Duby R, Olson E . Akt1/protein kinase B enhances transcriptional reprogramming of fibroblasts to functional cardiomyocytes. Proc Natl Acad Sci U S A. 2015; 112(38):11864-9. PMC: 4586885. DOI: 10.1073/pnas.1516237112. View

Banani S, Lee H, Hyman A, Rosen M . Biomolecular condensates: organizers of cellular biochemistry. Nat Rev Mol Cell Biol. 2017; 18(5):285-298. PMC: 7434221. DOI: 10.1038/nrm.2017.7. View

Christoforou N, Chellappan M, Adler A, Kirkton R, Wu T, Addis R . Transcription factors MYOCD, SRF, Mesp1 and SMARCD3 enhance the cardio-inducing effect of GATA4, TBX5, and MEF2C during direct cellular reprogramming. PLoS One. 2013; 8(5):e63577. PMC: 3660533. DOI: 10.1371/journal.pone.0063577. View

Savastano A, de Opakua A, Rankovic M, Zweckstetter M . Nucleocapsid protein of SARS-CoV-2 phase separates into RNA-rich polymerase-containing condensates. Nat Commun. 2020; 11(1):6041. PMC: 7699647. DOI: 10.1038/s41467-020-19843-1. View

Wang A, Bertos N, Vezmar M, Pelletier N, CROSATO M, Heng H . HDAC4, a human histone deacetylase related to yeast HDA1, is a transcriptional corepressor. Mol Cell Biol. 1999; 19(11):7816-27. PMC: 84849. DOI: 10.1128/MCB.19.11.7816. View

Grethe S, Porn-Ares M . p38 MAPK regulates phosphorylation of Bad via PP2A-dependent suppression of the MEK1/2-ERK1/2 survival pathway in TNF-alpha induced endothelial apoptosis. Cell Signal. 2005; 18(4):531-40. DOI: 10.1016/j.cellsig.2005.05.023. View

Hirai H, Kikyo N . Inhibitors of suppressive histone modification promote direct reprogramming of fibroblasts to cardiomyocyte-like cells. Cardiovasc Res. 2014; 102(1):188-90. PMC: 3958621. DOI: 10.1093/cvr/cvu023. View

Wang L, Liu Z, Yin C, Zhou Y, Liu J, Qian L . Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5. J Vis Exp. 2015; (105). PMC: 4692711. DOI: 10.3791/53426. View

10.

Zhou Y, Wang L, Vaseghi H, Liu Z, Lu R, Alimohamadi S . Bmi1 Is a Key Epigenetic Barrier to Direct Cardiac Reprogramming. Cell Stem Cell. 2016; 18(3):382-95. PMC: 4779178. DOI: 10.1016/j.stem.2016.02.003. View

11.

Wang B, Yang H, Liu Y, Jelinek T, Zhang L, Ruoslahti E . Isolation of high-affinity peptide antagonists of 14-3-3 proteins by phage display. Biochemistry. 1999; 38(38):12499-504. DOI: 10.1021/bi991353h. View

12.

Singh A, Ye M, Bucur O, Zhu S, Tanya Santos M, Rabinovitz I . Protein phosphatase 2A reactivates FOXO3a through a dynamic interplay with 14-3-3 and AKT. Mol Biol Cell. 2010; 21(6):1140-52. PMC: 2836964. DOI: 10.1091/mbc.e09-09-0795. View

13.

Wang L, Liu Z, Yin C, Asfour H, Chen O, Li Y . Stoichiometry of Gata4, Mef2c, and Tbx5 influences the efficiency and quality of induced cardiac myocyte reprogramming. Circ Res. 2014; 116(2):237-44. PMC: 4299697. DOI: 10.1161/CIRCRESAHA.116.305547. View

14.

Elbaum-Garfinkle S, Kim Y, Szczepaniak K, Chen C, Eckmann C, Myong S . The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics. Proc Natl Acad Sci U S A. 2015; 112(23):7189-94. PMC: 4466716. DOI: 10.1073/pnas.1504822112. View

15.

Wang Y, Shibasaki F, Mizuno K . Calcium signal-induced cofilin dephosphorylation is mediated by Slingshot via calcineurin. J Biol Chem. 2005; 280(13):12683-9. DOI: 10.1074/jbc.M411494200. View

16.

Zhu X, Chen L, Bai Q, Ma L, Li D, Zhang J . PP2A-AMPKα-HSF1 axis regulates the metal-inducible expression of HSPs and ROS clearance. Cell Signal. 2014; 26(4):825-32. DOI: 10.1016/j.cellsig.2014.01.002. View

17.

Soding J, Zwicker D, Sohrabi-Jahromi S, Boehning M, Kirschbaum J . Mechanisms for Active Regulation of Biomolecular Condensates. Trends Cell Biol. 2019; 30(1):4-14. DOI: 10.1016/j.tcb.2019.10.006. View

18.

Hornbeck P, Kornhauser J, Latham V, Murray B, Nandhikonda V, Nord A . 15 years of PhosphoSitePlus®: integrating post-translationally modified sites, disease variants and isoforms. Nucleic Acids Res. 2018; 47(D1):D433-D441. PMC: 6324072. DOI: 10.1093/nar/gky1159. View

19.

Lu J, McKinsey T, Nicol R, Olson E . Signal-dependent activation of the MEF2 transcription factor by dissociation from histone deacetylases. Proc Natl Acad Sci U S A. 2000; 97(8):4070-5. PMC: 18151. DOI: 10.1073/pnas.080064097. View

20.

Sabari B, DallAgnese A, Boija A, Klein I, Coffey E, Shrinivas K . Coactivator condensation at super-enhancers links phase separation and gene control. Science. 2018; 361(6400). PMC: 6092193. DOI: 10.1126/science.aar3958. View