Catching global interactions in vivo

Yi Qiu,Suming Huang

doi:10.1186/s13578-017-0177-z

Abstract

Histone proteins and transcription factors (TFs) play critical roles in gene transcription and development of multicellular organisms. Although antibody mediated protein isolation couple with mass spectrometry approach has been a standard method to identify TF interacting partners and characterize their functional molecular complexes, it becomes urge to develop a robust method to functional characterize how these transcription factors act during biological process in the post-human genome project era. Here, Dr. Zhao and his colleagues in the National Heart, Lung, and Blood Institute of NIH develop a sensitive and robust strategy to globally identify and characterize in vivo protein–protein interactions termed bait protein–protein interaction-sequencing (bPPI-seq) (Zhang et al. in Cell Res doi:10.1038/cr.2017.112, 2017). As a proof-of-principle, they demonstrated that genome-wide interacting partners of histone variant H2A.Z are mainly involved in transcriptional regulation which is distinct from the interacting proteins of canonical histone H2A. Thus, their results suggest that bPPI-seq can be widely used to globally characterize protein complexes especially transcription factor interacting partners and molecular networks formed.

Highlights

Transcription is a key mechanism underlying the control of gene activities and cell identity during animal development and disease
3 UF Health Cancer Center, University of Florida College of Medicine, Gainesville, FL 32610, USA Full list of author information is available at the end of the article volumes of starting materials and it may not be feasible for studies involved in certain cell lineages or disease samples
In bPPI-seq, the bait protein is fused to N-terminal green fluorescent protein (GFP) moiety while whole genome endogenous genes are randomly tagged by exon trapping using an enhanced retroviral mutagen vector containing cDNA of C-terminal half of GFP moiety and a doxycycline-inducible promoter driven splicing donor to induce a hybrid transcript of GFPC and in frame fused endogenous genes that allow expression of GFPC fusion proteins

Summary

Introduction

Transcription is a key mechanism underlying the control of gene activities and cell identity during animal development and disease. 3 UF Health Cancer Center, University of Florida College of Medicine, Gainesville, FL 32610, USA Full list of author information is available at the end of the article volumes of starting materials and it may not be feasible for studies involved in certain cell lineages or disease samples. As a transcription factor, their interactions required a physiological chromatin environment.

Results

Conclusion