title: C3 exoenzyme impairs cell proliferation and apoptosis by altering the activity of transcription factors.
abstract: C3 exoenzyme from C. botulinum is an ADP-ribosyltransferase that inactivates selectively RhoA, B, and C by coupling an ADP-ribose moiety. Rho-GTPases are involved in various cellular processes, such as regulation of actin cytoskeleton, cell proliferation, and apoptosis. Previous studies of our group with the murine hippocampal cell line HT22 revealed a C3-mediated inhibition of cell proliferation after 48 h and a prevention of serum-starved cells from apoptosis. For both effects, alterations of various signaling pathways are already known, including also changes on the transcriptional level. Investigations on the transcriptional activity in HT22 cells treated with C3 for 48 h identified five out of 48 transcription factors namely Sp1, ATF2, E2F-1, CBF, and Stat6 with a significantly regulated activity. For validation of identified transcription factors, studies on the protein level of certain target genes were performed. Western blot analyses exhibited an enhanced abundance of Sp1 target genes p21 and COX-2 as well as an increase in phosphorylation of c-Jun. In contrast, the level of p53 and apoptosis-inducing GADD153, a target gene of ATF2, was decreased. Our results reveal that C3 regulates the transcriptional activity of Sp1 and ATF2 resulting downstream in an altered protein abundance of various target genes. As the affected proteins are involved in the regulation of cell proliferation and apoptosis, thus the C3-mediated anti-proliferative and anti-apoptotic effects are consequences of the Rho-dependent alterations of the activity of certain transcriptional factors.
authors.affiliation: Institute of Toxicology, Hannover Medical School, Straße 1, D-30625, Hannover, Germany. vonelsner.leonie@mh-hannover.de.
authors.forename: Leonie
authors.initials: L
authors.lastname: von Elsner
authors.affiliation: Institute of Toxicology, Hannover Medical School, Straße 1, D-30625, Hannover, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Hagemann
authors.affiliation: Institute of Toxicology, Hannover Medical School, Straße 1, D-30625, Hannover, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.affiliation: Institute of Toxicology, Hannover Medical School, Straße 1, D-30625, Hannover, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Rohrbeck
mesh_terms: D036002:ADP Ribose Transferases / Q000494:pharmacology*; D051698:Activating Transcription Factor 2 / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001905:Botulinum Toxins / Q000494:pharmacology*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D006624:Hippocampus / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D010766:Phosphorylation; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013997:Time Factors; D051742:Transcription Factor CHOP / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: D051698:Activating Transcription Factor 2; C497106:Atf2 protein, mouse; C493957:Cdkn1a protein, mouse; D050759:Cyclin-Dependent Kinase Inhibitor p21; C497249:DDIT3 protein, human; D016755:Proto-Oncogene Proteins c-jun; D016329:Sp1 Transcription Factor; C495901:TP53 protein, human; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D051742:Transcription Factor CHOP; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D048051:p38 Mitogen-Activated Protein Kinases; D001905:Botulinum Toxins; D020741:rho GTP-Binding Proteins
keywords: Apoptosis; C3 exoenzyme; Proliferation; RhoA; Transcription factors