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Zhihong Ma,Zhanjun Lv,Xiu-Fang Wang,Xiangyang Jing,Jianjun Cheng 한국유전학회 2014 Genes & Genomics Vol.36 No.4
Cis-regulatory elements are regions of DNAthat regulate the expression of genes located on that samemolecule of DNA. Though these elements are important forgene expression regulation, the functions of cis elementsremain largely unknown. To explore the mechanisms ofgene activation by short sequence enhancers, we examinedthe enhancer activity of short sequence DNA and itsderived sequences using a GFP expression system. Wefound that AA sequence (50-GTGAAATAAATGCAAATAAAGT) induced strong GFP gene expression, while7pieA (50-GTGAAAAAAATGCAAAAAAAGT) did not. We mutated the five T bases of the AA sequence to A, C orG. Our findings indicated that sequences retaining the 7thand/or 17th Ts possessed strong enhancer activity. RT-PCRand RNA synthesis inhibition analysis using actinomycin Drevealed that the enhanced GFP gene expression inducedby the AA sequence occurred at the transcriptional level. To determine whether the AA sequence formed a secondarystructure via atypical complementation, PAGE methodwas used, and the results showed that the AA sequenceformed a secondary structure. Our results support previousevidence that AATAAA is an important composition of ciselements (enhancer/promoter), and suggest that the formationof an unstable stem-loop structure via atypicalcomplementation may be a new mechanism of enhanceractivity.