Immunoprecipitation of 11 ZN fingers Domains, CTCF and BORIS to Multifunctional Y-box DNA/RNA-binding factor, YB-1 in Glioma-RGBM

Abstract

CCCTC binding factor (CTCF) is a unique highly conserved and ubiquitously expressed 11 zinc finger (ZF) transcriptional factor with multiple target site. It is able to bind to various target sequences to perform different regulatory roles and the binding is through the combination of different ZF domains. On the other hand, BORIS (Brother of the Regulator of Imprinted Sites), which is expressed only in the testis and certain cancer cell lines is homology to CTCF 11 ZF domains. Since both transcriptional factors share the same ZF domains, hence there is a possibility for both to bind to the same target sequences. Hence, the aim of this study is to determine the in vivo interaction of CTCF and BORIS to YB-1 in the laboratory established Glioma-RGBM cell line. The protein-protein interaction between CTCF/YB-1 and BORIS/YB-1 were discovered using Co-immunoprecipitation (CO-IP) technique through reciprocal experiment in using RGBM total cell lysate. Results showed that both CTCF and BORIS were able to interact with YB-1 in RGBM cell line. To the best of our knowledge, this is the first finding demonstrating the ability of BORIS and YB-1 to form a complex in vivo

Key words: Immunoprecipitation; CTCF/BORIS/YB-1; Transcription factor