HEXIM1 Diffusion in the Nucleus Is Regulated by Its Interactions with Both 7SK and P-TEFb

Biophys J. 2019 Nov 5;117(9):1615-1625. doi: 10.1016/j.bpj.2019.09.019. Epub 2019 Sep 20.

Abstract

How nuclear proteins diffuse and find their targets remains a key question in the transcription field. Dynamic proteins in the nucleus are classically subdiffusive and undergo anomalous diffusion, yet the underlying physical mechanisms are still debated. In this study, we explore the contribution of interactions to the generation of anomalous diffusion by the means of fluorescence spectroscopy and simulation. Using interaction-deficient mutants, our study indicates that HEXIM1 interactions with both 7SK RNA and positive transcription elongation factor b are critical for HEXIM1 subdiffusion and thus provides evidence of the effects of protein-RNA interaction on molecular diffusion. Numerical simulations allowed us to establish that the proportions of distinct oligomeric HEXIM1 subpopulations define the apparent anomaly parameter of the whole population. Slight changes in the proportions of these oligomers can lead to significant shifts in the diffusive features and recapitulate the modifications observed in cells with the various interaction-deficient mutants. By combining simulations and experiments, our work opens new prospects in which the anomaly α coefficient in diffusion becomes a helpful tool to infer alterations in molecular interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Nucleus / metabolism*
  • Computer Simulation
  • Diffusion
  • Humans
  • Models, Molecular
  • Positive Transcriptional Elongation Factor B / metabolism*
  • Protein Binding
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • RNA-Binding Proteins / metabolism*
  • Spectrometry, Fluorescence
  • Transcription Factors / metabolism*

Substances

  • HEXIM1 protein, human
  • RNA, Long Noncoding
  • RNA-Binding Proteins
  • Transcription Factors
  • long non-coding RNA 7SK, human
  • Positive Transcriptional Elongation Factor B