title: The intermediate filament protein vimentin is essential for axonotrophic effects of Clostridium botulinum C3 exoenzyme.
abstract: The type III intermediate filament protein vimentin was recently identified to mediate binding and uptake of Clostridium botulinum C3 exoenzyme (C3bot) in two cell lines. Here, we used primary neuronal cultures from vimentin knockout (Vim-/-) mice to study the impact of vimentin on axonal growth and internalization of C3bot. In contrast to wild type, vimentin knockout neurons were insensitive to C3bot. Application of extracellular vimentin to Vim-/-neurons completely restored the growth-promoting effects of C3bot. In line with this uptake of C3bot into Vim-/-neurons was strongly decreased resulting in reduced ADP-ribosylation of RhoA and B as detected by an antibody recognizing selectively ADP-ribosylated RhoA/B. Again, uptake of C3bot into Vim-/-neurons was rescued by addition of extracellular vimentin. In addition, in purified embryonic stem cell-derived motor neurons that are devoid of glial cells C3bot elicited axonotrophic effects confining neuronal vimentin as a binding partner. Primary neuronal cultures from vimentin knockout (KO) mice were used to study the impact of vimentin on axonal growth and internalization of C3bot. In contrast to wild type, vimentin knockout neurons were insensitive to the axonotrophic effects of C3bot. Application of extracellular vimentin (recombinant vimentin) to vimentin KO neurons completely restored the growth-promoting effects of C3bot. In line with this uptake of C3bot into vimentin KO neurons was strongly decreased resulting in reduced ADP-ribosylation of RhoA and B as detected by an antibody recognizing selectively ADP-ribosylated RhoA/B.
authors.affiliation: Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Andrej
authors.initials: A
authors.lastname: Adolf
authors.affiliation: Department of Pharmacology, Medical School, University of Ioannina, Ioannina, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Leondaritis
authors.affiliation: Institute of Toxicology, Hannover Medical School (MHH), Hannover, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Rohrbeck
authors.affiliation: Institute of Biochemistry & Neuro Cure Cluster of Excellence, Charité - Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Britta Johanna
authors.initials: BJ
authors.lastname: Eickholt
authors.affiliation: Institute of Toxicology, Hannover Medical School (MHH), Hannover, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.affiliation: Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Gudrun
authors.initials: G
authors.lastname: Ahnert-Hilger
authors.affiliation: Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Berlin, Germany. markus.hoeltje@charite.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Höltje
mesh_terms: D036002:ADP Ribose Transferases / Q000494:pharmacology*; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D001369:Axons / Q000187:drug effects*; D001905:Botulinum Toxins / Q000494:pharmacology*; D002460:Cell Line; D005838:Genotype; D051379:Mice; D057507:Mice, 129 Strain; D018345:Mice, Knockout; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism; D058953:Neural Stem Cells / Q000378:metabolism; D061251:Primary Cell Culture; D014746:Vimentin / Q000235:genetics / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D020742:rhoA GTP-Binding Protein; D020743:rhoB GTP-Binding Protein / Q000378:metabolism
chemical_list: D014746:Vimentin; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; C526380:RhoA protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D020743:rhoB GTP-Binding Protein
keywords: C3 exoenzyme; axon outgrowth; vimentin