title: Systems analysis identifies an essential role for SHANK-associated RH domain-interacting protein (SHARPIN) in macrophage Toll-like receptor 2 (TLR2) responses.
abstract: Precise control of the innate immune response is essential to ensure host defense against infection while avoiding inflammatory disease. Systems-level analyses of Toll-like receptor (TLR)-stimulated macrophages suggested that SHANK-associated RH domain-interacting protein (SHARPIN) might play a role in the TLR pathway. This hypothesis was supported by the observation that macrophages derived from chronic proliferative dermatitis mutation (cpdm) mice, which harbor a spontaneous null mutation in the Sharpin gene, exhibited impaired IL-12 production in response to TLR activation. Systems biology approaches were used to define the SHARPIN-regulated networks. Promoter analysis identified NF-κB and AP-1 as candidate transcription factors downstream of SHARPIN, and network analysis suggested selective attenuation of these pathways. We found that the effects of SHARPIN deficiency on the TLR2-induced transcriptome were strikingly correlated with the effects of the recently described hypomorphic L153P/panr2 point mutation in Ikbkg [NF-κB Essential Modulator (NEMO)], suggesting that SHARPIN and NEMO interact. We confirmed this interaction by co-immunoprecipitation analysis and furthermore found it to be abrogated by panr2. NEMO-dependent signaling was affected by SHARPIN deficiency in a manner similar to the panr2 mutation, including impaired p105 and ERK phosphorylation and p65 nuclear localization. Interestingly, SHARPIN deficiency had no effect on IκBα degradation and on p38 and JNK phosphorylation. Taken together, these results demonstrate that SHARPIN is an essential adaptor downstream of the branch point defined by the panr2 mutation in NEMO.
authors.affiliation: Seattle Biomedical Research Institute, Seattle, WA 98109, USA.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Zak
authors.forename: Frank
authors.initials: F
authors.lastname: Schmitz
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Gold
authors.forename: Alan H
authors.initials: AH
authors.lastname: Diercks
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Peschon
authors.forename: Joe S
authors.initials: JS
authors.lastname: Valvo
authors.forename: Antti
authors.initials: A
authors.lastname: Niemistö
authors.forename: Irina
authors.initials: I
authors.lastname: Podolsky
authors.forename: Shannon G
authors.initials: SG
authors.lastname: Fallen
authors.forename: Rosa
authors.initials: R
authors.lastname: Suen
authors.forename: Tetyana
authors.initials: T
authors.lastname: Stolyar
authors.forename: Carrie D
authors.initials: CD
authors.lastname: Johnson
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Kennedy
authors.forename: M Kristina
authors.initials: MK
authors.lastname: Hamilton
authors.forename: Owen M
authors.initials: OM
authors.lastname: Siggs
authors.forename: Bruce
authors.initials: B
authors.lastname: Beutler
authors.forename: Alan
authors.initials: A
authors.lastname: Aderem
mesh_terms: D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D007113:Immunity, Innate / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism; D025941:Protein Interaction Mapping; D015398:Signal Transduction; D013597:Systems Analysis; D049490:Systems Biology; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D047908:Intracellular Signaling Peptides and Proteins; C514981:NEMO protein, mouse; D016328:NF-kappa B; C554944:Sipl1 protein, mouse; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2; D018808:Transcription Factor AP-1