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Chinese Journal of Hematology ; (12): 325-328, 2008.
Article Dans Chinois | WPRIM | ID: wpr-240016

Résumé

<p><b>OBJECTIVE</b>To identify the interaction partners of a new splicing product of LMO2 gene (LMO2-C), and study its function in K562 cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>LMO2-C can bind endogenous GATA1 and LDB1 protein in K562 cells and down regulates the expression of GPA.</p>


Sujets)
Humains , Protéines adaptatrices de la transduction du signal , Protéines de liaison à l'ADN , Génétique , Métabolisme , Facteur de transcription GATA-1 , Métabolisme , Cellules K562 , Protéines à domaine LIM , Protéines de liaison au maltose , Métalloprotéines , Génétique , Métabolisme , Protéines de liaison périplasmiques , Protéines proto-oncogènes , Épissage des ARN , Facteurs de transcription , Métabolisme , Techniques de double hybride
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