Identification of interaction partners and function analysis of new splicing product of human LMO2 gene.
- Author:
Wei YUAN
1
;
Shuang YANG
;
Wei SUN
;
Jun DU
;
Chun-Li ZHAI
;
Zhao-Qi WANG
;
Tian-Hui ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Adaptor Proteins, Signal Transducing; DNA-Binding Proteins; genetics; metabolism; GATA1 Transcription Factor; metabolism; Humans; K562 Cells; LIM Domain Proteins; Maltose-Binding Proteins; Metalloproteins; genetics; metabolism; Periplasmic Binding Proteins; Proto-Oncogene Proteins; RNA Splicing; Transcription Factors; metabolism; Two-Hybrid System Techniques
- From: Chinese Journal of Hematology 2008;29(5):325-328
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the interaction partners of a new splicing product of LMO2 gene (LMO2-C), and study its function in K562 cells.
METHODSMaltose 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.
RESULTSMBP-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.
CONCLUSIONLMO2-C can bind endogenous GATA1 and LDB1 protein in K562 cells and down regulates the expression of GPA.
