Simultaneous visualization of multiple protein interactions in living cells using multicolor fluorescence complementation analysis.
The specificity of biological regulatory mechanisms relies on selective interactions between different proteins in different cell types and in response to different extracellular signals. We describe a bimolecular fluorescence complementation (BiFC) approach for the simultaneous visualization of multiple protein interactions in the same cell. This approach is based on complementation between fragments of fluorescent proteins with different spectral characteristics. We have identified 12 bimolecular fluorescent complexes that correspond to 7 different spectral classes. Bimolecular complex formation between fragments of different fluorescent proteins did not differentially affect the dimerization efficiency of the bZIP domains of Fos and Jun or the subcellular sites of interactions between these domains. Multicolor BiFC enables visualization of interactions between different proteins in the same cell and comparison of the efficiencies of complex formation with alternative interaction partners.
Hu
Chang-Deng
CD
Howard Hughes Medical Institute and Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109-0650, USA.
Kerppola
Tom K
TK
D000818:Animals; D050976:Basic-Leucine Zipper Transcription Factors; D019556:COS Cells; D002522:Chlorocebus aethiops; D003116:Color; D004268:DNA-Binding Proteins / Q000032:analysis / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D005456:Fluorescent Dyes; D051790:G-Box Binding Factors; D008856:Microscopy, Fluorescence / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000145:classification / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000032:analysis / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D016335:Zinc Fingers
D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D051790:G-Box Binding Factors; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors