Screening and detecting proteins interacting with the N terminal of MINT

    ZHOU Peng1£¬ LI Junª²Feng1£¬ QIN Hong1£¬HAN Hua1,2

    1Department of Medical Genetics & Developmental Biology, School of Basic Medicine, Xi¬ðan 710033, China, 2Stem Cell Center of Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038, China

    ¡¾Abstract¡¿ AIM: To study the mechanism of the repression of MINT, a 3576aa protein in nuclear matrix, and to further clarify the regulation of the pathway of Notch, yeast two hybrid assay was used to screen molecules which can interact with MINTª²F1(amino acids 1ª²365). METHODS: The bait vector was constructed by inserting Mintª²F1 into pGBKT7 to screen the cDNA library of human lymph node. The positive clones were restrictively digested and sequenced for further analysis. RESULTS: Seventyª²three clones were obtained after 6¡Á107 clones were yielded by the four kinds of nutrition limitation and ¦Âª²galactosidase assays. Twelve positive clones were obtained after restriction of positive clones. Sequencing of the clones showed that there were 3 significative molecules: small nuclear ribonucleoprotein polypeptide G (SNRPG), Vav oncogene and microspherule protein 1 (MCRS1). CONCLUSION: Analysing with the 3 molecules obtained from the human cDNA library, we can see that the function of MINT may be complex, it may participate in the splicing of RNA by affecting the spliceosomal small nuclear ribonucleoproteins (snRNPs), or regulate the cellular signal transduction pathway by interacting with some signal transduction factors, such as Vav1 and RBPª²J¦Ê. It may also affect the size and shape of the nucleolus through MCRS1. ......