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General Information
Name
RTK-Ras-ERK_Interactome
Species
D. melanogaster
Result type
FlyBase ID
FBlc0000669
Project
Data Provider
Title
386 high-confidence interactions involving components of the RTK-Ras-Erk signaling pathway.
Status
Current
Accessions
    Biosample Source
    Overview
    Strain
    Stage
    Sex
    Tissue isolated
    Other tissues studied
    Cell component
    Key genes
    Sample preparation

    cDNAs for 15 components of the RTK-Ras-ERK pathway were cloned into into pMK33-CTAP and stable S2R+ cell lines were generated. Cells were induced overnight with 140uM copper. At least two biological replicates were performed for each bait and condition.

    Biosamples analyzed by this result (1)
    Biosample
    Type
    Title
    D. melanogaster, S2R+ cell line, stably transfected with bait protein, source for RTK-Ras-ERK pathway affinity purifications.
    Data Analyzed
    Assay methods
    Key genes
    (bait_protein)
    Protocol

    Cells were lysed as described in Veraksa et al., 2005 (FBrf0183909) and in-solution tandem affinity purification was performed as described in Siomi and Siomi, 2005 (FBrf0188010).

    Mode of Assay

    Purified proteins were reduced with DTT, alkylated with iodoacetamide, trypsin digested, cleaned of buffer and debris by 'ZipTip' and dried to concentrate peptides. Peptides were analysed by LC-MS/MS. MS/MS spectra were matched to peptides using SEQUEST and a composite database of all translated D. melanogaster open reading frames in FlyBase (r5.4).

    Raw Data Analyzed (1)
    Assay / Reagent collection
    Type
    Title
    Coaffinity purification of interacting proteins and identification by mass spectrometry, S2R+ cells, RTK-Ras-ERK pathway components.
    Processed Data Analyzed (0)
    Result
    Type
    Title
    Analysis
    Methods
    Reference Genome
    Reference Annotation
    Data analysis

    From 15 baits, 5009 unfiltered interactions among 1188 proteins were identified. SAINT analysis was used to assign confidence scores (PMID: 21131968). 'Tag-only' purifications were used as negative controls. Using a SAINT cutoff of 0.83 and a false discovery rate of 7.2%, a high confidence, filtered network of 386 interactions among 249 proteins was generated.

    Comments
    Associated Data
    Size
    Associated interactions
    386 Interactions
    Additional Information
    Synonyms and Secondary IDs (4)
    Reported As
    Symbol Synonym
    RTK-Ras-ERK interaction network
    RTK-Ras-ERK_Interactome
    Name Synonyms
    386 high-confidence interactions involving components of the RTK-Ras-Erk signaling pathway.
    Secondary FlyBase IDs
      References (2)