c-Jun controls histone modifications, NF-B recruitment and RNA polymerase II function to activate the ccl2 gene

Institute of Pharmacology, Medical School Hannover, Carl-Neuberg Strasse 1, D-30625, Hannover, Germany; Institute of Molecular Pathology, Vienna, A-1030, Austria

^* To whom correspondence should be addressed. Email: Michael.Kracht{at}pharma.med.uni-giessen.de.

	Abstract

IL-1-induced mRNA expression of ccl2 (also called MCP-1), a prototypic highly regulated inflammatory gene, is severely suppressed in cells lacking c-Jun or JNK1/JNK2 genes and is only partially restored in cells expressing a c-Jun(SS63/73AA) mutant. By chromatin immunoprecipitation we identified three c-Jun-binding sites located in the far 5' region, close to the transcriptional start site and in the far 3' region of murine and human ccl2 genes. Mutational analysis revealed that the latter two contribute to ccl2 transcription in response to IL-1, or to ectopically expressed c-Jun-ATF-2 dimers. Further experiments comparing wild type and c-Jun-deficient cells revealed that c-Jun regulates ser10 phosphorylation of histone H3, acetylation of histones H3 and H4 and recruitment of HDAC3, NF-B subunits and RNA polymerase II across the ccl2 locus. c-Jun also co-immunoprecipitated with p65 NF-B and HDAC3. Based on DNA microarray analysis, c-Jun was required for full expression of 133 out of 162 IL-1-induced genes. For inflammatory genes, these data support an activator function of c-Jun that is executed by multiple mechanisms, including phosphorylation-dependent interaction with p65 NF-B and HDAC3 at the level of chromatin.