Yuan Wei, Yang Shuang, Sun Wei, Du Jun, Zhai Chun-Li, Wang Zhao-Qi, Zhu Tian-Hui
Laboratory of Molecular Genetics, Medical College, Nankai University, Tianjin 300071, China.
Zhonghua Xue Ye Xue Za Zhi. 2008 May;29(5):325-8.
To identify the interaction partners of a new splicing product of LMO2 gene (LMO2-C), and study its function in K562 cells.
Maltose binding protein (MBP) pull down and mammalian two-hybrid assay (MTHA) were used to identify the interaction partners of LMO2-C in K562 cells. Semiquantitative RT-PCR was used to detect the expression of hematopoietic specific gene glycoprotein (GPA) in K562 cells.
MBP-LMO2-C fusion protein was expressed and purified in soluble form successfully. Endogenous GATA1 and LDB1 proteins were confirmed to bind to LMO2-C by MBP pull down analysis. The MTHA also showed that LMO2-C had comparable binding affinities to LDB1 with LMO2-L, and over expression of LMO2-C prevented LMO2-L from binding to LDB1, the inhibition rate being (81.13 +/- 0.68)%. RT-PCR results showed that the expression level of GPA was reduced [(51.00 +/- 1.58)%] in K562 cells while LMO2-C overexpressed.
LMO2-C can bind endogenous GATA1 and LDB1 protein in K562 cells and down regulates the expression of GPA.
