TAILIEUCHUNG - Báo cáo khoa học: Cloning and functional analysis of 5¢-upstream region of the Pokemon gene

Pokemon, the POK erythroid myeloid ontogenic factor, not only regulates the expression of many genes, but also plays an important role in cell tumorigenesis. To investigate the molecular mechanism regulating expres-sion of the Pokemongene in humans, its 5¢-upstream region was cloned and analyzed. | ễFEBS Journal Cloning and functional analysis of 5 -upstream region of the Pokemon gene Yutao Yang Xiaowei Zhou Xudong Zhu Chuanfu Zhang Zhixin Yang Long Xu and Peitang Huang Laboratory of Protein Engineering Beijing Institute of Biotechnology China Keywords DNA decoy element mutation Pokemon promoter Correspondence P. Huang Laboratory of Protein Engineering Beijing Institute of Biotechnology Beijing 100071 China Fax Tel 86 10 6381 0272 E-mail amms832@ Received 15 October 2007 revised 12 February 2008 accepted 18 February 2008 doi Pokemon the POK erythroid myeloid ontogenic factor not only regulates the expression of many genes but also plays an important role in cell tumorigenesis. To investigate the molecular mechanism regulating expression of the Pokemon gene in humans its 5 -upstream region was cloned and analyzed. Transient analysis revealed that the Pokemon promoter is constitutive. Deletion analysis and a DNA decoy assay indicated that the NEG-U and NEG-D elements were involved in negative regulation of the Pokemon promoter whereas the POS-D element was mainly responsible for its strong activity. Electrophoretic mobility shift assays suggested that the NEG-U NEG-D and POS-D elements were specifically bound by the nuclear extract from A549 cells in vitro. Mutation analysis demonstrated that cooperation of the NEG-U and NEG-D elements led to negative regulation of the Pokemon promoter. Moreover the NEG-U and NEG-D elements needed to be an appropriate distance apart in the Pokemon promoter in order to cooperate. Taken together our results elucidate the mechanism underlying the regulation of Pokemon gene transcription and also define a novel regulatory sequence that may be used to decrease expression of the Pokemon gene in cancer gene therapy. The poxvirus and zinc finger POZ domain formerly termed the broad complex tramtrack and bric-a-brac BTB domain was initially characterized in the Drosophila proteins broad complex .

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