[Identification of interaction partners and function analysis of new splicing product of human LMO2 gene].
OBJECTIVE
To identify the interaction partners of a new splicing product of LMO2 gene (LMO2-C), and study its function in K562 cells.


METHODS
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.


RESULTS
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.


CONCLUSIONS
LMO2-C can bind endogenous GATA1 and LDB1 protein in K562 cells and down regulates the expression of GPA.
Laboratory of Molecular Genetics, Medical College, Nankai University, Tianjin 300071, China.
Wei
W
Yuan
Shuang
S
Yang
Wei
W
Sun
Jun
J
Du
Chun-Li
CL
Zhai
Zhao-Qi
ZQ
Wang
Tian-Hui
TH
Zhu
D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D050982:GATA1 Transcription Factor / Q000378:metabolism; D006801:Humans; D020014:K562 Cells; D060588:LIM Domain Proteins; D058619:Maltose-Binding Proteins; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism*; D033902:Periplasmic Binding Proteins; D011518:Proto-Oncogene Proteins; D012326:RNA Splicing; D014157:Transcription Factors / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C486312:LDB1 protein, human; D060588:LIM Domain Proteins; C085117:LMO2 protein, human; D058619:Maltose-Binding Proteins; D008667:Metalloproteins; D033902:Periplasmic Binding Proteins; D011518:Proto-Oncogene Proteins; D014157:Transcription Factors