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PMID: 11160903 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transcriptional activation of the small GTPase gene rhoB by genotoxic stress is regulated via a CCAAT element.

Nucleic acids research ·Vol. 29 ·No. 3 ·2001-02-01 ·Pages 792-8

Fritz G, Kaina B

Abstract

The gene encoding the Ras-related GTPase RhoB-specific is immediate-early inducible by genotoxic treatments. Regulation of transcriptional activation of rhoB is still unclear. Here we show that cells lacking either p53 or c-Fos are not different from wild-type cells with respect to the level of rhoB induction upon UV irradiation, indicating that these transcription factors are not crucial for stimulation of rhoB mRNA expression. Extracts from UV-irradiated and non-irradiated cells revealed similar DNA-binding activities to a 0.17 kb rhoB promoter fragment harboring the functional element(s) necessary for stimulation of rhoB by UV light. By means of immunoprecipitation we found that an ATF-2-specific antibody co-precipitates the (32)P-labeled 0.17 kb rhoB fragment, whereas an anti-AP1 antibody did not. Since no consensus sequence for binding of ATF-2 is present within the rhoB promoter, ATF-2 is likely to be associated with another factor that binds to the minimal promoter. Deletion analysis and site-directed mutagenesis of the 0.17 kb rhoB fragment revealed a CCAAT box to be an essential requirement for stimulation of rhoB by UV light and methyl methanesulfonate. Moreover, immunoprecipitation experiments showed that the CCAAT-binding factor NF-YA is complexed with ATF-2. Overall, the data strongly indicate that transcriptional activation of the rhoB gene by genotoxic stress is regulated via a CCAAT box and that interaction of CCAAT-binding factor and ATF-2 triggers the stress-inducible expression of rhoB.

MeSH Terms
3T3 Cells Animals Base Sequence Binding Sites CCAAT-Binding Factor/genetics,metabolism CCAAT-Enhancer-Binding Proteins/genetics,metabolism DNA/genetics,metabolism Mice Molecular Sequence Data Mutation Oligonucleotides/genetics,metabolism Promoter Regions, Genetic/genetics Protein Binding/radiation effects RNA, Messenger/genetics,metabolism,radiation effects Response Elements/genetics Transcription Factors/metabolism Transcriptional Activation/radiation effects Ultraviolet Rays rhoB GTP-Binding Protein/genetics
Chemicals
CCAAT-Binding Factor CCAAT-Enhancer-Binding Proteins Nfya protein, mouse Oligonucleotides RNA, Messenger Transcription Factors DNA rhoB GTP-Binding Protein
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fritz G
Division of Applied Toxicology, Institute of Toxicology, University of Mainz, Obere Zahlbacher Strasse 67, D-55131 Mainz, Germany. fritz@mail.uni-mainz.de
Kaina B
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/29.3.792
Email
fritz@mail.uni-mainz.de
Published
2001-02-01
Pages
792-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC30383
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/29.3.792 PMC Full Text View journal details
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