Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation

Chang-Deng Hu; Yurii Chinenov; Tom K Kerppola

doi:10.1016/s1097-2765(02)00496-3

Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation

Mol Cell. 2002 Apr;9(4):789-98. doi: 10.1016/s1097-2765(02)00496-3.

Authors

Chang-Deng Hu¹, Yurii Chinenov, Tom K Kerppola

Affiliation

¹ Howard Hughes Medical Institute and Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

PMID: 11983170
DOI: 10.1016/s1097-2765(02)00496-3

Abstract

Networks of protein interactions coordinate cellular functions. We describe a bimolecular fluorescence complementation (BiFC) assay for determination of the locations of protein interactions in living cells. This approach is based on complementation between two nonfluorescent fragments of the yellow fluorescent protein (YFP) when they are brought together by interactions between proteins fused to each fragment. BiFC analysis was used to investigate interactions among bZIP and Rel family transcription factors. Regions outside the bZIP domains determined the locations of bZIP protein interactions. The subcellular sites of protein interactions were regulated by signaling. Cross-family interactions between bZIP and Rel proteins affected their subcellular localization and modulated transcription activation. These results attest to the general applicability of the BiFC assay for studies of protein interactions.

MeSH terms

3T3 Cells
Alternative Splicing
Animals
Bacterial Proteins / chemistry*
COS Cells
Cell Compartmentation
Chlorocebus aethiops
DNA-Binding Proteins / chemistry*
Dimerization
Fluorescence
HeLa Cells
Humans
I-kappa B Proteins*
Leucine Zippers*
Ligases / chemistry*
Luminescent Proteins / chemistry*
Macromolecular Substances
Mice
Microscopy, Fluorescence
NF-KappaB Inhibitor alpha
NF-kappa B / chemistry*
Peptide Fragments / chemistry
Protein Interaction Mapping / methods*
Protein Structure, Tertiary
Protein Transport
Proto-Oncogene Proteins c-fos / chemistry*
Proto-Oncogene Proteins c-jun / chemistry*
Recombinant Fusion Proteins / chemistry
Sequence Deletion
Subcellular Fractions / chemistry*
Transcription, Genetic
Transfection

Substances

Bacterial Proteins
DNA-Binding Proteins
I-kappa B Proteins
Luminescent Proteins
Macromolecular Substances
NF-kappa B
NFKBIA protein, human
Nfkbia protein, mouse
Peptide Fragments
Proto-Oncogene Proteins c-fos
Proto-Oncogene Proteins c-jun
Recombinant Fusion Proteins
yellow fluorescent protein, Bacteria
NF-KappaB Inhibitor alpha
Ligases
guanosine 3',5'-polyphosphate synthetases