Gadd45a protects against UV irradiation-induced skin tumors, and promotes apoptosis and stress signaling via MAPK and p53.

Skin cancer is the most frequent form of malignancy in the world, and UV radiation is the primary environmental carcinogen responsible for its development. Herein we demonstrate that Gadd45a is a critical factor protecting the epidermis against UV radiation-induced tumorigenesis by promoting damaged keratinocytes to undergo apoptosis and/or cell cycle arrest, two crucial events that prevent the expansion of mutant or deregulated cells. Whereas Gadd45a has been implicated in cell cycle arrest, apoptosis, and DNA repair, to determine the physiological function of endogenous Gadd45a after genotoxic stress, the skin of Gadd45a-null mice was targeted with UV radiation. We report that Gadd45a induces apoptosis and cell cycle arrest by maintaining p38 and c-JNK MAPK activation in keratinocytes. The absence of Gadd45a results in loss of sustained p38/JNK MAPK activity beyond 15-30 min after UV radiation that leads to inadequate p53 activation and loss of normal activation of G(1) and G(2) checkpoints. Moreover, loss of Gadd45a dramatically reduces UV-induced apoptotic keratinocytes, "sunburn cells." Consequently, Gadd45a-null mice are more prone to tumors relative to wild-type mice. Therefore, we conclude that Gadd45a, like p53, is a key component protecting skin against UV-induced tumors.

[1]  J. Saurat,et al.  Upregulation of connexin 26 between keratinocytes of psoriatic lesions. , 1998, The Journal of investigative dermatology.

[2]  P M Steinert,et al.  Expression of murine epidermal differentiation markers is tightly regulated by restricted extracellular calcium concentrations in vitro , 1989, The Journal of cell biology.

[3]  J. Simon,et al.  A role for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[4]  J. Lord,et al.  Serine/threonine protein kinases and apoptosis. , 2000, Experimental cell research.

[5]  M. Karin,et al.  NF-kappa B activation by ultraviolet light not dependent on a nuclear signal. , 1993, Science.

[6]  T. Jacks,et al.  Sunburn and p53 in the onset of skin cancer , 1994, Nature.

[7]  Wei Zhang,et al.  CR6: A third member in the MyD118 and Gadd45 gene family which functions in negative growth control , 1999, Oncogene.

[8]  L. Donehower,et al.  Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours , 1992, Nature.

[9]  Michael Karin,et al.  Ultraviolet Light and Osmotic Stress: Activation of the JNK Cascade Through Multiple Growth Factor and Cytokine Receptors , 1996, Science.

[10]  D. Bar-Sagi,et al.  Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. , 2000, Science.

[11]  M. Cobb,et al.  Mitogen-activated protein kinase pathways. , 1997, Current opinion in cell biology.

[12]  P. O'Connor,et al.  Direct interaction of Gadd45 with PCNA and evidence for competitive interaction of Gadd45 and p21Waf1/Cip1 with PCNA. , 1995, Oncogene.

[13]  G. Stark,et al.  Regulation of the G2/M transition by p53 , 2001, Oncogene.

[14]  J. Fargnoli,et al.  Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents , 1989, Molecular and cellular biology.

[15]  P. Hanawalt,et al.  p53-Mediated DNA Repair Responses to UV Radiation: Studies of Mouse Cells Lacking p53, p21, and/orgadd45 Genes , 2000, Molecular and Cellular Biology.

[16]  D. Brash,et al.  Skin precancer. , 1998, Cancer surveys.

[17]  P. O'Connor,et al.  Abrogation of p53 function affects gadd gene responses to DNA base-damaging agents and starvation. , 1996, DNA and cell biology.

[18]  M. Yaffe,et al.  MAP kinase pathways activated by stress: the p38 MAPK pathway. , 2000, Critical care medicine.

[19]  Kohei Miyazono,et al.  Positive and negative regulation of TGF-β signaling , 2000 .

[20]  H. K. Sluss,et al.  Selective interaction of JNK protein kinase isoforms with transcription factors. , 1996, The EMBO journal.

[21]  M. Karin,et al.  Stress-induced JNK Activation Is Independent of Gadd45 Induction* , 1999, The Journal of Biological Chemistry.

[22]  A. Fornace,et al.  Dimethylbenzanthracene carcinogenesis in Gadd45a-null mice is associated with decreased DNA repair and increased mutation frequency. , 2001, Cancer research.

[23]  K. Sakaguchi,et al.  Phosphorylation of human p53 by p38 kinase coordinates N‐terminal phosphorylation and apoptosis in response to UV radiation , 1999, The EMBO journal.

[24]  U. Mohr,et al.  International Classification of Rodent Tumors. The Mouse , 2001, Springer Berlin Heidelberg.

[25]  R. Flavell,et al.  GADD45gamma mediates the activation of the p38 and JNK MAP kinase pathways and cytokine production in effector TH1 cells. , 2001, Immunity.

[26]  J. Cadet,et al.  Effects of UV and visible radiations on cellular DNA. , 2001, Current problems in dermatology.

[27]  T. Myers,et al.  Gadd45, a p53-Responsive Stress Protein, Modifies DNA Accessibility on Damaged Chromatin , 1999, Molecular and Cellular Biology.

[28]  Kazushige Yamaguchi,et al.  Clinicopathological significance of abnormalities in Gadd45 expression and its relationship to p53 in human pancreatic cancer. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[29]  E. Appella,et al.  Initiation of a G2/M checkpoint after ultraviolet radiation requires p38 kinase , 2001, Nature.

[30]  R. Pfundt,et al.  In situ demonstration of phosphorylated c‐jun and p38 MAP kinase in epidermal keratinocytes following ultraviolet B irradiation of human skin , 2001, The Journal of pathology.

[31]  M. Skolnick,et al.  A cell cycle regulator potentially involved in genesis of many tumor types. , 1994, Science.

[32]  M. Pincus,et al.  Jun NH2-Terminal Kinase Phosphorylation of p53 on Thr-81 Is Important for p53 Stabilization and Transcriptional Activities in Response to Stress , 2001, Molecular and Cellular Biology.

[33]  X. Wang,et al.  Association with Cdc2 and inhibition of Cdc2/Cyclin B1 kinase activity by the p53-regulated protein Gadd45 , 1999, Oncogene.

[34]  H. Saito,et al.  A Family of Stress-Inducible GADD45-like Proteins Mediate Activation of the Stress-Responsive MTK1/MEKK4 MAPKKK , 1998, Cell.

[35]  Mutsuhiro Takekawa,et al.  Regulation of MTK1/MEKK4 Kinase Activity by Its N-Terminal Autoinhibitory Domain and GADD45 Binding , 2002, Molecular and Cellular Biology.

[36]  L. Donehower,et al.  Susceptibility to radiation-carcinogenesis and accumulation of chromosomal breakage in p53 deficient mice. , 1994, Oncogene.

[37]  J. Avruch,et al.  Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. , 2001, Physiological reviews.

[38]  R. Brent,et al.  Mxi2, a mitogen-activated protein kinase that recognizes and phosphorylates Max protein. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[39]  M. Karin,et al.  The mammalian ultraviolet response is triggered by activation of src tyrosine kinases , 1992, Cell.

[40]  A. Fornace,et al.  Cyclobutane pyrimidine dimers in UV-DNA induce release of soluble mediators that activate the human immunodeficiency virus promoter. , 1993, The Journal of investigative dermatology.

[41]  C. Prives,et al.  The p53 pathway , 1999, The Journal of pathology.

[42]  P. O'Connor,et al.  Interaction of the p53-regulated protein Gadd45 with proliferating cell nuclear antigen. , 1994, Science.

[43]  K. Kohn,et al.  The gadd and MyD genes define a novel set of mammalian genes encoding acidic proteins that synergistically suppress cell growth , 1994, Molecular and cellular biology.

[44]  P. Hanawalt,et al.  p 53-Mediated DNA Repair Responses to UV Radiation : Studies of Mouse Cells Lacking p 53 , p 21 , and / or gadd 45 Genes , 2000 .

[45]  William F. Morgan,et al.  Genomic instability in Gadd45a-deficient mice , 1999, Nature Genetics.

[46]  M. Gorospe,et al.  gadd45 Is Not Required for Activation of c-Jun N-terminal Kinase or p38 during Acute Stress* , 1999, The Journal of Biological Chemistry.

[47]  Francesca Zazzeroni,et al.  Induction of gadd45β by NF-κB downregulates pro-apoptotic JNK signalling , 2001, Nature.

[48]  Peter Greaves,et al.  I – Integumentary System , 2000 .

[49]  Kendall A. Smith,et al.  Cytokine response gene 6 induces p21 and regulates both cell growth and arrest , 1999, Oncogene.

[50]  D. Livingston,et al.  Binding and modulation of p53 by p300/CBP coactivators , 1997, Nature.

[51]  C. Prives,et al.  p53: puzzle and paradigm. , 1996, Genes & development.

[52]  T. Hirano,et al.  A Novel Oncostatin M-inducible Gene OIG37 Forms a Gene Family with MyD118 and GADD45 and Negatively Regulates Cell Growth* , 1999, The Journal of Biological Chemistry.

[53]  A. Quinn Ultraviolet radiation and skin carcinogenesis. , 1997, British journal of hospital medicine.

[54]  J. Avruch,et al.  Protein kinase cascades activated by stress and inflammatory cytokines , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[55]  K. Hashimoto Regulation of keratinocyte function by growth factors. , 2000, Journal of dermatological science.

[56]  Xiaozhong Wang,et al.  Stress-Induced Phosphorylation and Activation of the Transcription Factor CHOP (GADD153) by p38 MAP Kinase , 1996, Science.

[57]  D. Brash,et al.  Ultraviolet radiation induced signature mutations in photocarcinogenesis. , 1999, The journal of investigative dermatology. Symposium proceedings.

[58]  D. McConkey,et al.  Temporal events in skin injury and the early adaptive responses in ultraviolet-irradiated mouse skin. , 2000, The American journal of pathology.

[59]  Albert J. Fornace,et al.  Amplification of PPM1D in human tumors abrogates p53 tumor-suppressor activity , 2002, Nature Genetics.

[60]  Y. Ip,et al.  Signal transduction by the c-Jun N-terminal kinase (JNK)--from inflammation to development. , 1998, Current opinion in cell biology.

[61]  J. Bartek,et al.  Pathways governing G1/S transition and their response to DNA damage , 2001, FEBS letters.

[62]  D. Liebermann,et al.  Sequence and expression of a cDNA encoding MyD118: a novel myeloid differentiation primary response gene induced by multiple cytokines. , 1991, Oncogene.

[63]  K. Scharffetter-Kochanek,et al.  UV-induced oxidative stress and photoaging. , 2001, Current problems in dermatology.

[64]  A. Prescott,et al.  Gadd45 is a nuclear cell cycle regulated protein which interacts with p21Cip1. , 1995, Oncogene.

[65]  L. Zon,et al.  Cdc42Hs, but Not Rac1, Inhibits Serum-stimulated Cell Cycle Progression at G1/S through a Mechanism Requiring p38/RK* , 1997, The Journal of Biological Chemistry.

[66]  M. Reiss,et al.  Functional evidence for an extracellular calcium receptor mechanism triggering tyrosine kinase activation associated with mouse keratinocyte differentiation. , 1994, The Journal of biological chemistry.

[67]  K. Holbrook,et al.  Potassium mediation of calcium-induced terminal differentiation of epidermal cells in culture. , 1983, The Journal of investigative dermatology.

[68]  C. Beadling,et al.  Isolation of interleukin 2-induced immediate-early genes. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[69]  Karen Holbrook,et al.  Calcium regulation of growth and differentiation of mouse epidermal cells in culture , 1980, Cell.

[70]  E. Martín-Blanco p38 MAPK signalling cascades: ancient roles and new functions , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[71]  A. Sarasin,et al.  The molecular pathways of ultraviolet-induced carcinogenesis. , 1999, Mutation research.

[72]  F. Christians,et al.  Induction of GADD45 and JNK/SAPK-Dependent Apoptosis following Inducible Expression of BRCA1 , 1999, Cell.

[73]  K. Isobe,et al.  Peroxynitrite induces GADD34, 45, and 153 VIA p38 MAPK in human neuroblastoma SH-SY5Y cells. , 2001, Free radical biology & medicine.

[74]  K. Miyazono,et al.  Positive and negative regulation of TGF-beta signaling. , 2000, Journal of cell science.

[75]  Sally A Amundson,et al.  Roles for p53 in growth arrest and apoptosis: putting on the brakes after genotoxic stress , 1998, Oncogene.

[76]  C. Sutter,et al.  Carcinogen-specific mutational pattern in the p53 gene in ultraviolet B radiation-induced squamous cell carcinomas of mouse skin. , 1992, Cancer research.

[77]  Quinn Ag Ultraviolet radiation and skin carcinogenesis. , 1997 .

[78]  V. Tron,et al.  Induction of squamous cell carcinoma in p53-deficient mice after ultraviolet irradiation. , 1998, The Journal of investigative dermatology.

[79]  R. Weichselbaum,et al.  Activation of p38 Mitogen-activated Protein Kinase by c-Abl-dependent and -independent Mechanisms* , 1996, The Journal of Biological Chemistry.

[80]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.