Bimolecular fluorescence complementation; lighting-up tau-tau interaction in living cells

PLoS One. 2013 Dec 2;8(12):e81682. doi: 10.1371/journal.pone.0081682. eCollection 2013.

Abstract

Abnormal tau aggregation is a pathological hallmark of many neurodegenerative disorders and it is becoming apparent that soluble tau aggregates play a key role in neurodegeneration and memory impairment. Despite this pathological importance, there is currently no single method that allows monitoring soluble tau species in living cells. In this regard, we developed a cell-based sensor that visualizes tau self-assembly. By introducing bimolecular fluorescence complementation (BiFC) technique to tau, we were able to achieve spatial and temporal resolution of tau-tau interactions in a range of states, from soluble dimers to large aggregates. Under basal conditions, tau-BiFC cells exhibited little fluorescence intensity, implying that the majority of tau molecules exist as monomers. Upon chemically induced tau hyperphosphorylation, BiFC fluorescence greatly increased, indicating an increased level of tau-tau interactions. As an indicator of tau assembly, our BiFC sensor would be a useful tool for investigating tau pathology.

MeSH terms

  • Cell Survival
  • Fluorescent Dyes / metabolism*
  • HEK293 Cells
  • Humans
  • Microtubules / metabolism
  • Models, Molecular
  • Molecular Imaging / methods*
  • Phosphorylation
  • Protein Binding
  • Protein Multimerization
  • Protein Structure, Secondary
  • tau Proteins / chemistry
  • tau Proteins / metabolism*

Substances

  • Fluorescent Dyes
  • tau Proteins

Grants and funding

This work was supported by an intramural funding from Korea Institute of Science and Technology (2E23870 and 2E24138, the Korea government (MSIP, number 2007-00559), and the National Research Foundation of Korea individual scientist supporting program (NRF-2012R1A1A2004980). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.