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Functional Dissection of an Enhancer-Like Element Located within the Second Intron of the Human U2AF1L4 Gene

D. A. Didych, N. A. Smirnov, E. S. Kotova, S. B. Akopov*, L. G. Nikolaev, and E. D. Sverdlov

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia; fax: (495) 330-6538; E-mail: akser@ibch.ru

* To whom correspondence should be addressed.

Received December 29, 2010; Revision received March 21, 2011
A detailed functional and evolutionary analysis of an enhancer element of the human genome (enhancer 12) located in the second intron of the U2AF1L4 gene, which we identified earlier, is presented. Overlapping fragments of the studied genome region were analyzed for enhancer activity, and the site responsible for the activity of this element was identified using transient transfections of HeLa cells. Comparison of the enhancer 12 sequence with orthologous sequences from seven primate species revealed the existence of evolutionarily conserved sequences within this element. One of the identified conservative regions is likely responsible for the enhancer activity and is able to specifically interact in vitro with proteins of HeLa cell nuclear extract. The ability of orthologous primate sequences to compete with enhancer 12 for binding with HeLa cell nuclear extract proteins and to enhance the activity of the reporter gene in transient transfection of HeLa cells is demonstrated.
KEY WORDS: human genome, regulation of gene expression, functional analysis, enhancer

DOI: 10.1134/S0006297911080116