Eya2 Promotes Cell Cycle Progression by Regulating DNA Damage Response During Vertebrate Limb Regeneration

Overview

Journal Elife

Specialty Biology

Date 2020 Mar 7

PMID 32142407

Citations 15

Authors

Konstantinos Sousounis

Donald M Bryant

Jose Martinez Fernandez

Samuel S Eddy

Stephanie L Tsai

Gregory C Gundberg

Jihee Han

Katharine Courtemanche

Michael Levin

Jessica L Whited

Affiliations

Soon will be listed here.

Abstract

How salamanders accomplish progenitor cell proliferation while faithfully maintaining genomic integrity and regenerative potential remains elusive. Here we found an innate DNA damage response mechanism that is evident during blastema proliferation (early- to late-bud) and studied its role during tissue regeneration by ablating the function of one of its components, Eyes absent 2. In mutant axolotls, we found that DNA damage signaling through the H2AX histone variant was deregulated, especially within the proliferating progenitors during limb regeneration. Ultimately, cell cycle progression was impaired at the G1/S and G2/M transitions and regeneration rate was reduced. Similar data were acquired using acute pharmacological inhibition of the Eya2 phosphatase activity and the DNA damage checkpoint kinases Chk1 and Chk2 in wild-type axolotls. Together, our data indicate that highly-regenerative animals employ a robust DNA damage response pathway which involves regulation of H2AX phosphorylation via Eya2 to facilitate proper cell cycle progression upon injury.

Citing Articles

BMP signaling promotes zebrafish heart regeneration via alleviation of replication stress.

Vasudevarao M, Posadas Pena D, Ihle M, Bongiovanni C, Maity P, Geissler D Nat Commun. 2025; 16(1):1708.

PMID: 39962064 PMC: 11832743. DOI: 10.1038/s41467-025-56993-6.

The In Vitro Enhancement of Retinal Cell Viability via mA and mC RNA Methylation-Mediated Changes in the Levels of Heme Oxygenase (HO-1) and DNA Damage Repair Molecules Using a 50 Hz Sinusoidal Electromagnetic Field (EMF).

Betlej G, Bator E, Koziorowska A, Koziorowski M, Rzeszutek I Int J Mol Sci. 2025; 25(24.

PMID: 39769368 PMC: 11677922. DOI: 10.3390/ijms252413606.

Negative cell cycle regulation by calcineurin is necessary for proper beta cell regeneration in zebrafish.

Massoz L, Bergemann D, Lavergne A, Reynders C, Desiront C, Goossens C Elife. 2024; 12.

PMID: 39383064 PMC: 11464004. DOI: 10.7554/eLife.88813.

Amputation Triggers Long-Range Epidermal Permeability Changes in Evolutionarily Distant Regenerative Organisms.

Dooling K, Kim R, Kim E, Chen E, Abouelela A, Tajer B bioRxiv. 2024; .

PMID: 39257748 PMC: 11383696. DOI: 10.1101/2024.08.29.610385.

DNA Damage, Genome Stability, and Adaptation: A Question of Chance or Necessity?.

Herrick J Genes (Basel). 2024; 15(4).

PMID: 38674454 PMC: 11049855. DOI: 10.3390/genes15040520.

References

McCusker C, Diaz-Castillo C, Sosnik J, Phan A, Gardiner D . Cartilage and bone cells do not participate in skeletal regeneration in Ambystoma mexicanum limbs. Dev Biol. 2016; 416(1):26-33. DOI: 10.1016/j.ydbio.2016.05.032. View

Nakamura A, Rao V, Pommier Y, Bonner W . The complexity of phosphorylated H2AX foci formation and DNA repair assembly at DNA double-strand breaks. Cell Cycle. 2010; 9(2):389-97. PMC: 3086803. DOI: 10.4161/cc.9.2.10475. View

Smith J, Tho L, Xu N, Gillespie D . The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Adv Cancer Res. 2010; 108:73-112. DOI: 10.1016/B978-0-12-380888-2.00003-0. View

Liu Y, Long Y, Wang S, Li Y, Zhang J . Phosphorylation of H2A.X positively regulates DNA damage response and is linked to cancer progression. FEBS J. 2016; 283(24):4462-4473. DOI: 10.1111/febs.13951. View

Smith J, Timoshevskaya N, Timoshevskiy V, Keinath M, Hardy D, Voss S . A chromosome-scale assembly of the axolotl genome. Genome Res. 2019; 29(2):317-324. PMC: 6360810. DOI: 10.1101/gr.241901.118. View

Brown J, Eykelenboom J, Lowndes N . Co-mutation of histone H2AX S139A with Y142A rescues Y142A-induced ionising radiation sensitivity. FEBS Open Bio. 2013; 2:313-7. PMC: 3678120. DOI: 10.1016/j.fob.2012.09.008. View

Elewa A, Wang H, Talavera-Lopez C, Joven A, Brito G, Kumar A . Reading and editing the Pleurodeles waltl genome reveals novel features of tetrapod regeneration. Nat Commun. 2017; 8(1):2286. PMC: 5741667. DOI: 10.1038/s41467-017-01964-9. View

Malumbres M, Barbacid M . Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009; 9(3):153-66. DOI: 10.1038/nrc2602. View

Villiard E, Brinkmann H, Moiseeva O, Mallette F, Ferbeyre G, Roy S . Urodele p53 tolerates amino acid changes found in p53 variants linked to human cancer. BMC Evol Biol. 2007; 7:180. PMC: 2072957. DOI: 10.1186/1471-2148-7-180. View

10.

Pimentel H, Bray N, Puente S, Melsted P, Pachter L . Differential analysis of RNA-seq incorporating quantification uncertainty. Nat Methods. 2017; 14(7):687-690. DOI: 10.1038/nmeth.4324. View

11.

Balic A, Adams D, Mina M . Prx1 and Prx2 cooperatively regulate the morphogenesis of the medial region of the mandibular process. Dev Dyn. 2009; 238(10):2599-613. PMC: 2806060. DOI: 10.1002/dvdy.22092. View

12.

Jemc J, Rebay I . The eyes absent family of phosphotyrosine phosphatases: properties and roles in developmental regulation of transcription. Annu Rev Biochem. 2007; 76:513-38. DOI: 10.1146/annurev.biochem.76.052705.164916. View

13.

Krishnan N, Jeong D, Jung S, Ryu S, Xiao A, Allis C . Dephosphorylation of the C-terminal tyrosyl residue of the DNA damage-related histone H2A.X is mediated by the protein phosphatase eyes absent. J Biol Chem. 2009; 284(24):16066-16070. PMC: 2713548. DOI: 10.1074/jbc.C900032200. View

14.

Wood R, Mitchell M, Sgouros J, Lindahl T . Human DNA repair genes. Science. 2001; 291(5507):1284-9. DOI: 10.1126/science.1056154. View

15.

Bedelbaeva K, Snyder A, Gourevitch D, Clark L, Zhang X, Leferovich J . Lack of p21 expression links cell cycle control and appendage regeneration in mice. Proc Natl Acad Sci U S A. 2010; 107(13):5845-50. PMC: 2851923. DOI: 10.1073/pnas.1000830107. View

16.

Szklarczyk D, Franceschini A, Wyder S, Forslund K, Heller D, Huerta-Cepas J . STRING v10: protein-protein interaction networks, integrated over the tree of life. Nucleic Acids Res. 2014; 43(Database issue):D447-52. PMC: 4383874. DOI: 10.1093/nar/gku1003. View

17.

Gong D, Ferrell Jr J . The roles of cyclin A2, B1, and B2 in early and late mitotic events. Mol Biol Cell. 2010; 21(18):3149-61. PMC: 2938381. DOI: 10.1091/mbc.E10-05-0393. View

18.

Huen M, Chen J . The DNA damage response pathways: at the crossroad of protein modifications. Cell Res. 2007; 18(1):8-16. DOI: 10.1038/cr.2007.109. View

19.

Whited J, Tsai S, Beier K, White J, Piekarski N, Hanken J . Pseudotyped retroviruses for infecting axolotl in vivo and in vitro. Development. 2013; 140(5):1137-46. PMC: 3583047. DOI: 10.1242/dev.087734. View

20.

Darnet S, Dragalzew A, Amaral D, Sousa J, Thompson A, Cass A . Deep evolutionary origin of limb and fin regeneration. Proc Natl Acad Sci U S A. 2019; 116(30):15106-15115. PMC: 6660751. DOI: 10.1073/pnas.1900475116. View