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UV Radiation-induced Impairment of Cellular Morphology and Motility is Enhanced by DUSP3/VHR Loss and FAK Activation

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Abstract

DUSP3 is a phosphatase expressed and active in several tissues that dephosphorylates tyrosine residues in many regulatory proteins of cellular activities such as proliferation, survival, and cell death. Recently, two new independent functions were assigned to this enzyme: dephosphorylation of focal adhesion kinase (FAK) and regulation of nucleotide-excision repair (NER) pathway. Genotoxic stress by UV radiation is known to affect cell morphology, adhesion, and migration for affecting, for example, the Rho GTPases that regulate actin cytoskeleton. This work investigated the intersection of DUSP3 function, XPA protein activity, and UV toxicity by examining cell migration, FAK, and SRC kinase phosphorylation status, in addition to cell morphology, in fibroblast cells proficient (MRC-5) or deficient (XPA) of the NER pathway. DUSP3 loss reduced cell migration of normal cells, which was stimulated by the genotoxic stress, effects evidenced in presence of serum mitogenic stimulus. However, NER-deficient cells migration response was the opposite since DUSP3 loss increased migration, especially after cells being exposed to UV stress. The levels of pFAK(Y397) peaked 15 min and 1 h after UV radiation in normal cells, but only slightly increased in repair-deficient cells. However, the DUSP3 knockdown strongly raised pFAK(Y397) levels in both cells, but especially in XPA cells as supported by the higher SRC activity. These effects impacted on the dynamics of actin-based structures formation, such as stress fibres, apparently dependent on DUSP3 and DNA-repair (NER) proficiency of the cells. Altogether our findings suggest this dual-phosphatase is bridging gaps between the complex regulation of cell morphology, motility, and genomic stability.

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Acknowledgements

The authors thank Prof. Bianca S. Zingales and the technician Marcelo Nunes Silva, from the Institute of Chemistry—University of São Paulo, for equipment usage and reagents exchange. The authors are grateful for the technical services of Izaura Nobuko Toma and Benedita de Oliveira from the Laboratory of Biomolecular Systems Signalling. The authors also thank Prof. Alexandre Bruni Cardoso from the Institute of Chemistry—University of São Paulo for helping with Leica microscope imaging and software analyses.

Funding

This work was primarily supported by Sao Paulo Research Foundation—FAPESP (Grants Nos. 2015/03983-0 and 2018/01753-6) and the Brazilian National Research Council—CNPQ (Grant No. 402230/2016-7). L.C.R. was founded by PNPD fellowship from the Ministry of Education through the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES (88887.136364/2017-00).

Author contributions

N.R.P. and L.C.R. performed most of the experimental study; F.L.F. supervised and designed this study; L.C.R. and F.L.F. analyzed and interpreted the data; and L.C.R. and F.L.F. wrote the paper. F.L.F. reviewed and reedited the manuscript.

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Authors and Affiliations

  1. Laboratory of Biomolecular Systems Signalling, Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil

    Nadine Ranieri Pereira, Lilian Cristina Russo & Fabio Luis Forti

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  1. Nadine Ranieri Pereira
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  2. Lilian Cristina Russo
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  3. Fabio Luis Forti
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Correspondence to Fabio Luis Forti.

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Pereira, N.R., Russo, L.C. & Forti, F.L. UV Radiation-induced Impairment of Cellular Morphology and Motility is Enhanced by DUSP3/VHR Loss and FAK Activation. Cell Biochem Biophys 79, 261–269 (2021). https://doi.org/10.1007/s12013-021-00966-1

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