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PMID: 10097101 Published · ppublish English Journal Article

GADD45 induction of a G2/M cell cycle checkpoint.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 7 ·1999-03-30 ·Pages 3706-11

Wang XW, Zhan Q, Coursen JD, Khan MA, Kontny HU, Yu L, Hollander MC, O'Connor PM, Fornace AJ, Harris CC

Abstract

G1/S and G2/M cell cycle checkpoints maintain genomic stability in eukaryotes in response to genotoxic stress. We report here both genetic and functional evidence of a Gadd45-mediated G2/M checkpoint in human and murine cells. Increased expression of Gadd45 via microinjection of an expression vector into primary human fibroblasts arrests the cells at the G2/M boundary with a phenotype of MPM2 immunopositivity, 4n DNA content and, in 15% of the cells, centrosome separation. The Gadd45-mediated G2/M arrest depends on wild-type p53, because no arrest was observed either in p53-null Li-Fraumeni fibroblasts or in normal fibroblasts coexpressed with p53 mutants. Increased expression of cyclin B1 and Cdc25C inhibited the Gadd45-mediated G2/M arrest in human fibroblasts, indicating that the mechanism of Gadd45-mediated G2/M checkpoint is at least in part through modulation of the activity of the G2-specific kinase, cyclin B1/p34(cdc2). Genetic and physiological evidence of a Gadd45-mediated G2/M checkpoint was obtained by using GADD45-deficient human or murine cells. Human cells with endogenous Gadd45 expression reduced by antisense GADD45 expression have an impaired G2/M checkpoint after exposure to either ultraviolet radiation or methyl methanesulfonate but are still able to undergo G2 arrest after ionizing radiation. Lymphocytes from gadd45-knockout mice (gadd45 -/-) also retained a G2/M checkpoint initiated by ionizing radiation and failed to arrest at G2/M after exposure to ultraviolet radiation. Therefore, the mammalian genome is protected by a multiplicity of G2/M checkpoints in response to specific types of DNA damage.

MeSH Terms
Animals Ataxia Telangiectasia Mutated Proteins Cell Cycle/physiology Cell Cycle Proteins Cell Line Colonic Neoplasms Cyclin-Dependent Kinase Inhibitor p21 Cyclins/genetics DNA Damage DNA-Binding Proteins Fibroblasts/cytology,physiology G2 Phase Genes, p53 Humans Intracellular Signaling Peptides and Proteins Lymphocytes/cytology,physiology Mice Mice, Inbred Strains Mitosis Protein Serine-Threonine Kinases Proteins/genetics,physiology Recombinant Proteins/metabolism Spleen/cytology Transfection Tumor Cells, Cultured Tumor Suppressor Proteins
Chemicals
CDKN1A protein, human Cdkn1a protein, mouse Cell Cycle Proteins Cyclin-Dependent Kinase Inhibitor p21 Cyclins DNA-Binding Proteins GADD45 protein Intracellular Signaling Peptides and Proteins Proteins Recombinant Proteins Tumor Suppressor Proteins ATM protein, human Ataxia Telangiectasia Mutated Proteins Atm protein, mouse Protein Serine-Threonine Kinases
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Wang X W
Laboratory of Human Carcinogenesis, Division of Basic Science, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Zhan Q
Coursen J D
Khan M A
Kontny H U
Yu L
Hollander M C
O'Connor P M
Fornace A J
Harris C C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.7.3706
Published
1999-03-30
Pages
3706-11
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC22358
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.7.3706 PMC Full Text View journal details
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