title: Threonine 149 phosphorylation enhances ΔFosB transcriptional activity to control psychomotor responses to cocaine.
abstract: Stable changes in neuronal gene expression have been studied as mediators of addicted states. Of particular interest is the transcription factor ΔFosB, a truncated and stable FosB gene product whose expression in nucleus accumbens (NAc), a key reward region, is induced by chronic exposure to virtually all drugs of abuse and regulates their psychomotor and rewarding effects. Phosphorylation at Ser(27) contributes to ΔFosB's stability and accumulation following repeated exposure to drugs, and our recent work demonstrates that the protein kinase CaMKIIα phosphorylates ΔFosB at Ser(27) and regulates its stability in vivo. Here, we identify two additional sites on ΔFosB that are phosphorylated in vitro by CaMKIIα, Thr(149) and Thr(180), and demonstrate their regulation in vivo by chronic cocaine. We show that phosphomimetic mutation of Thr(149) (T149D) dramatically increases AP-1 transcriptional activity while alanine mutation does not affect transcriptional activity when compared with wild-type (WT) ΔFosB. Using in vivo viral-mediated gene transfer of ΔFosB-T149D or ΔFosB-T149A in mouse NAc, we determined that overexpression of ΔFosB-T149D in NAc leads to greater locomotor activity in response to an initial low dose of cocaine than does WT ΔFosB, while overexpression of ΔFosB-T149A does not produce the psychomotor sensitization to chronic low-dose cocaine seen after overexpression of WT ΔFosB and abrogates the sensitization seen in control animals at higher cocaine doses. We further demonstrate that mutation of Thr(149) does not affect the stability of ΔFosB overexpressed in mouse NAc, suggesting that the behavioral effects of these mutations are driven by their altered transcriptional properties.
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Hannah M
authors.initials: HM
authors.lastname: Cates
authors.affiliation: Department of Physiology, Michigan State University, Lansing, Michigan 48824.
authors.forename: Mackenzie
authors.initials: M
authors.lastname: Thibault
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Madeline
authors.initials: M
authors.lastname: Pfau
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Elizabeth
authors.identifier: http://orcid.org/0000-0002-2984-8705
authors.initials: E
authors.lastname: Heller
authors.affiliation: Department of Physiology, Michigan State University, Lansing, Michigan 48824.
authors.forename: Andrew
authors.identifier: http://orcid.org/0000-0002-7879-5996
authors.initials: A
authors.lastname: Eagle
authors.affiliation: Department of Physiology, Michigan State University, Lansing, Michigan 48824.
authors.forename: Paula
authors.initials: P
authors.lastname: Gajewski
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Rosemary
authors.initials: R
authors.lastname: Bagot
authors.affiliation: Department of Molecular Biophysics and Biochemistry/Keck Biotechnology Services, New Haven, Connecticut 06511, and.
authors.forename: Christopher
authors.initials: C
authors.lastname: Colangelo
authors.affiliation: Department of Molecular Biophysics and Biochemistry/Keck Biotechnology Services, New Haven, Connecticut 06511, and.
authors.forename: Thomas
authors.initials: T
authors.lastname: Abbott
authors.affiliation: Department of Pharmacology/Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, Texas 77555.
authors.forename: Gabby
authors.initials: G
authors.lastname: Rudenko
authors.affiliation: Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
authors.forename: Rachael
authors.initials: R
authors.lastname: Neve
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Nestler
authors.affiliation: Department of Physiology, Michigan State University, Lansing, Michigan 48824, robiso45@msu.edu.
authors.forename: Alfred J
authors.initials: AJ
authors.lastname: Robison
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D003042:Cocaine / Q000494:pharmacology*; D018765:Dopamine Uptake Inhibitors / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009447:Neuroblastoma / Q000473:pathology; D009714:Nucleus Accumbens / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics / Q000378:metabolism*; D011597:Psychomotor Performance / Q000187:drug effects*; D013912:Threonine / Q000235:genetics / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism
chemical_list: D018765:Dopamine Uptake Inhibitors; D016760:Proto-Oncogene Proteins c-fos; D018808:Transcription Factor AP-1; D049452:Green Fluorescent Proteins; D013912:Threonine; D000255:Adenosine Triphosphate; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D003042:Cocaine
keywords: CaMKII; accumbens; cocaine; phosphorylation; transcription; δFosB