Screening Differentiation Related GCN5 Recruitment Proteins during Differentiation of Mesenchymal Stem Cells into Cardiac Like Myocytes

Abstract: Screening and analysing the recruitment of protein complexes with GCN5 during the process of mesenchymal stem cells (MSCs) differentiating into cardiac myocytes have been designed to identify unknown proteins that were recruited by histone acetyltransferase GCN5. Immunofluorescence cytochemistry and real-time PCR analysis identify that MSCs can differentiate into cardiomyocyte-like cells after 5-azaC induction treatment. We then developed co-immunoprecipitation strategy to separate recruitment proteins with GCN5， and tandem mass spectrometry to identify recruitment proteins. Furthermore， we analyzed functions of GCN5 recruitment proteins from the degree of differentiation of cardiac. MSCs can express higher cardiac-specific genes GATA4 and MEF2C after 5-azaC induction treatment. GCN5 physically interacts with the following proteins which can be categorized as: (1) DNA-binding protein of Sp1/KLF; (2) transcriptional co-activators of PGC-1a and Rb1; (3) components of transcription elongation complex， and finally signaling pathway component of protein Akt. This experiment screens GCN5 recruitment proteins for regulation. We will further study differentiating signal transduction pathway of stem cells， as well as the specific biological target for intervention to improve the ratio of differentiation of stem cells on the basis of experiments.

CSTR: 32200.14.cjcb.2010.01.0009