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Journal of Biochemistry Advance Access originally published online on April 24, 2007
Journal of Biochemistry 2007 142(1):25-31; doi:10.1093/jb/mvm098
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© 2007 The Japanese Biochemical Society.

Studying on the 19-bp Palindrome Repeats in Human Cytomegalovirus Immediate Early Enhancer/Promoter Reveals their Diversity in Function for the Promoter Activity

Ping Shen1,3, Gang Niu1,3, Minghui Yao1,3, Haoyue Wang1 and Jian Fei1,2,4,*

1Model Organism Research Center and Laboratory of Molecular Cell Biology/Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; 2Shanghai Nan Fang Model Organism Research Center, Pu Dong, Shanghai 201203, China; 3Graduate School of the Chinese Academy of Sciences, Beijing 100049, China; and 4Model Organism Division, E-Institutes of the Shanghai Universities, Shanghai Jiao Tong University, Shanghai 200025, China

*To whom correspondence should be addressed. Jian Fei, No. 88, Cailun Road, Pu Dong, Shanghai 201203, China. Tel: +86 21 58951009, Fax: +8621 58951005. E-mail: jfei{at}sibs.ac.cn

Received April 11, 2007; Accepted April 11, 2007


  Abstract

The human cytomegalovirus major immediate early enhancer/promoter (HCMV MIEP), extending from –588 to +1 relative to the transcription start site, contains a series of reiterated cis-acting elements, such as 19-bp repeats, which occur four times in the enhancer/promoter region. However, it is still not clear whether these elements repeat just for backing up or they really execute various indispensable functions. We show here that these reiterated elements are functionally different from each other through serial deletion mutation and site-directed mutation. In addition, we also found that the CG-reverse in the first 19-bp repeat could improve the transcription activity of MIEP in HeLa cells and impair the protein-binding capacity in the EMSA assay. The expression feature of this mutated MIEP in transgenic mice further confirmed its stronger and more universal transcription activity in vivo.

Key Words: HCMV, transcription regulation, MIEP, P sites, transgenic mice

Abbreviations: HCMV, human cytomegalovirus; MIEP, major immediate early enhancer/promoter; P sites, 19-bp palindrome repeats; P1 site, the first 19-bp palindrome repeat relative to the transcription start site; 1mP MIEP, the mutated MIEP with a CG-reverse in the sequence of P1 site

The authors wish it to be known that, in their opinion, the first three authors should be regarded as joint First Authors.


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