Regulation of mouse GADD34 gene transcription after DNA damaging agent methylmethane sulfonate.
The GADD34 gene is transcriptionally induced by growth arrest and DNA damage. However, the mechanisms underlying the transcriptional regulation are still unclear. We analyzed the promoter of mouse GADD34 gene and the methylmethane sulfonate (MMS)-induced transcriptional regulation of this gene. By introducing genome mutants, which were linked to the luciferase reporter, into NIH3T3 cells, we defined a 100-bp fragment upstream of the transcriptional initiating site as the minimal promoter of the GADD34 gene. Subsequent study revealed that CRE-binding site located in this minimal promoter was critical for MMS-induced transcription of the GADD34 gene. In vitro binding experiments showed that phosphorylated c-Jun was contained in the CRE/DNA complex. Overexpression of the dominant negative form of c-Jun led to a decrease of MMS-responsive promoter activity. From these results, we conclude that the CRE site of the GADD34 promoter is indispensable to the MMS-responsive cis-element that c-Jun is the essential transcription factor for MMS-stimulated regulation of GADD34 gene expression and that the upstream signaling is dependent on JNK.
Haneda
Masataka
M
Department of Basic Gerontology, National Institute for Longevity Sciences, 36-3 Gengo, Morioka-cho, Obu, Aichi 474-8522, Japan.
Xiao
Hengyi
H
Hasegawa
Tadao
T
Kimura
Yuko
Y
Nakashima
Izumi
I
Isobe
Ken-Ichi
K
D051698:Activating Transcription Factor 2; D000818:Animals; D000943:Antigens, Differentiation; D001665:Binding Sites / Q000235:genetics; D018797:Cell Cycle Proteins; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D015151:Immunoblotting; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D048670:MAP Kinase Kinase 4; D008741:Methyl Methanesulfonate / Q000633:toxicity*; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation; D041681:NIH 3T3 Cells; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D054645:Protein Phosphatase 1; D011506:Proteins / Q000235:genetics*; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection
D051698:Activating Transcription Factor 2; D000943:Antigens, Differentiation; D018797:Cell Cycle Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D009153:Mutagens; D011506:Proteins; D016755:Proto-Oncogene Proteins c-jun; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D008741:Methyl Methanesulfonate; D008156:Luciferases; D048670:MAP Kinase Kinase 4; C482562:Map2k4 protein, mouse; D020929:Mitogen-Activated Protein Kinase Kinases; C513256:Ppp1r15a protein, mouse; D054645:Protein Phosphatase 1