Pull-Down assay between CTCF- N, ZF and C-terminal domains and YB-1 – N, CSD and C terminal domains in RGBM cell line.

Research Journal of Pharmaceutical, Biological and Chemical Sciences

article

Nur Natasha¹  Daruliza Kernain¹  Shaharum Shamsuddin.¹ 

[1]Institute for Research in Molecular Medicine

[2]Advanced Molecular Biology Laboratory, School of Health Sciences, Health Campus

关键词: Immunoprecipitation;    CTCF/BORIS/YB-1;    Transcription factor;   

DOI  :  10.33887/rjpbcs/2019.10.3.3

学科分类：工程和技术（综合）

来源: Research Journal of Pharmaceutical, Biological and Chemical Sciences

【 摘 要 】

CTCF is an 11-Zn finger transcription factor, involved in the regulation of transcription, insulator function,and the X-chromosome inactivation. CTCF is ubiquitously expressed and it is highly conserved between the species.Identification of proteins interacting with CTCF can help to understand the biological function of CTCF in the cell.Previously reported studies have identified numerous CTCF protein interacting partners and two of the proteinpartners were chosen for this study. The first protein was the transcription factor YB-1, a member of the Y-boxfamily and the second protein was the Large Subunit of RNA Polymerase II (LS Pol II), the principal enzyme fortranscription. The interaction of CTCF and YB-1 was characterized in the Glioma-RGBM cell line. The interactionbetween these two proteins was characterized through in vitro pull-down assay. From the results obtained, CTCF- ZF was the only domain binds with YB-1 CSD. The rest of domains did not show any interaction. Next, thesignificant of functional interaction between these two transcriptional factors was determined via mammaliantwo-hybrid system.The results showed a strong interaction between CTCF and YB-1 when both were co- transfected into RGBM cell line which proved the biological significant of these two proteins interaction in thecell.Thus the findings demonstrate, for the first time, the biological relevance of the CTCF/YB-1 interaction in theRGBM cell line.

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