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J. Biol. Chem., Vol. 281, Issue 34, 24623-24636, August 25, 2006

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Transcription of Human Zinc Finger ZNF268 Gene Requires an Intragenic Promoter Element^*

Ming-Xiong Guo¹, Di Wang¹, Huan-Jie Shao, Hong-Ling Qiu, Lu Xue, Zhou-Zhou Zhao, Chen-Gang Zhu, Yun-Bo Shi², and Wen-Xin Li³

From the State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China and Section on Molecular Morphogenesis, Laboratory of Gene Regulation and Development, NICHD, National Institutes of Health, Bethesda, Maryland 20892

Human ZNF268 gene is a typical Krüppel-associated box/C2H2 zinc finger gene whose homolog has been found only in higher mammals and not in lower mammals such as mouse. Its expression profiles have suggested that it plays a role in the differentiation of blood cells during early human embryonic development and the pathogenesis of leukemia. To gain additional insight into the molecular mechanisms controlling the expression of the ZNF268 gene and to provide the necessary tools for further genetic studies of leukemia, we have mapped the 5'-end of the human ZNF268 mRNA by reverse transcription-PCR and primer extension assays. We then cloned the 5'-flanking genomic DNA containing the putative ZNF268 gene promoter and analyzed its function in several different human and mouse tissue culture cell lines. Interestingly, our studies show that the ZNF268 gene lacks a typical eukaryotic promoter that is present upstream of the transcription start site and directs a basal level of transcription. Instead, the functional promoter requires an essential element that is located within the first exon of the gene. Deletion and mutational analysis reveals the requirement for a cAMP response-element-binding protein (CREB)-binding site within this element for promoter function. Gel mobility shift and chromatin immunoprecipitation assays confirm that CREB-2 binds to the site in vitro and in vivo. Furthermore, overexpression of CREB-2 enhances the promoter activity. These results demonstrate that the human ZNF268 gene promoter is atypical and requires an intragenic element located within the first exon that mediates the effect of CREB for its activity.

Received for publication, March 23, 2006 , and in revised form, June 20, 2006.

^* This work was supported in part by the National Natural Science Foundation of China Grant 30500266 and the Research Fund for the Doctoral Program of Higher Education Grant 20020486044. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² Supported by the Intramural Research Program of the NICHD, National Institutes of Health.

³ To whom correspondence should be addressed. Tel.: 86-27-68752831; Fax: 86-27-68752146; E-mail: Liwxlab{at}whu.edu.cn.

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