title: Hsc70 is a novel interactor of NF-kappaB p65 in living hippocampal neurons.
abstract: Signaling via NF-κB in neurons depends on complex formation with interactors such as dynein/dynactin motor complex and can be triggered by synaptic activation. However, so far a detailed interaction map for the neuronal NF-κB is missing. In this study we used mass spectrometry to identify novel interactors of NF-κB p65 within the brain. Hsc70 was identified as a novel neuronal interactor of NF-κB p65. In HEK293 cells, a direct physical interaction was shown by co-immunoprecipitation and verified via in situ proximity ligation in healthy rat neurons. Pharmacological blockade of Hsc70 by deoxyspergualin (DSG) strongly decreased nuclear translocation of NF-κB p65 and transcriptional activity shown by reporter gene assays in neurons after stimulation with glutamate. In addition, knock down of Hsc70 via siRNA significantly reduced neuronal NF-κB activity. Taken together these data provide evidence for Hsc70 as a novel neuronal interactor of NF-κB p65.
authors.affiliation: Cell Biology, University of Bielefeld, Bielefeld, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Klenke
authors.forename: Darius
authors.initials: D
authors.lastname: Widera
authors.forename: Thomas
authors.initials: T
authors.lastname: Engelen
authors.forename: Janine
authors.initials: J
authors.lastname: Müller
authors.forename: Thomas
authors.initials: T
authors.lastname: Noll
authors.forename: Karsten
authors.initials: K
authors.lastname: Niehaus
authors.forename: M Lienhard
authors.initials: ML
authors.lastname: Schmitz
authors.forename: Barbara
authors.initials: B
authors.lastname: Kaltschmidt
authors.forename: Christian
authors.initials: C
authors.lastname: Kaltschmidt
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D050883:HSC70 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology*; D006801:Humans; D051379:Mice; D009154:Mutation / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D034421:Sus scrofa; D013696:Temperature; D051996:Transcription Factor RelA / Q000378:metabolism*
chemical_list: D050883:HSC70 Heat-Shock Proteins; C494372:Hspa8 protein, mouse; C494373:Hspa8 protein, rat; D034741:RNA, Small Interfering; D051996:Transcription Factor RelA; D000255:Adenosine Triphosphate