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Originally published In Press as doi:10.1074/jbc.M209700200 on October 25, 2002

J. Biol. Chem., Vol. 277, Issue 51, 49700-49706, December 20, 2002
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Orientation-dependent Influence of an Intergenic Enhancer on the Promoter Activity of the Divergently Transcribed Mouse Shsp/alpha B-crystallin and Mkbp/HspB2 Genes*

Shivalingappa K. Swamynathan and Joram PiatigorskyDagger

From the Laboratory of Molecular and Developmental Biology, NEI, National Institutes of Health, Bethesda, Maryland 20892

The mouse Shsp/alpha B-crystallin and Mkbp/HspB2 genes are closely linked and divergently transcribed. In this study, we have analyzed the contribution of the intergenic enhancer to Shsp/alpha B-crystallin and Mkbp/HspB2 promoter activity using dual-reporter vectors in transient transfection and transgenic mouse experiments. Deletion of the enhancer reduced Shsp/alpha B-crystallin promoter activity by 30- and 93-fold and Mkbp/HspB2 promoter activity by 6- and 10-fold in transiently transfected mouse lens alpha -TN4 and myoblast C2C12 cells, respectively. Surprisingly, inversion of the enhancer reduced Shsp/alpha B-crystallin promoter activity by 17-fold, but did not affect Mkbp/HspB2 promoter activity in the transfected cells. In contrast, enhancer activity was orientation-independent in combination with a heterologous promoter in transfected cells. Transgenic mouse experiments established the orientation dependence and Shsp/alpha B-crystallin promoter preference of the intergenic enhancer in its native context. The orientation dependence and preferential effect of the Shsp/alpha B-crystallin enhancer on the Shsp/alpha B-crystallin promoter provide an example of adaptive changes in gene regulation accompanying the functional diversification of duplicated genes during evolution.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Laboratory of Molecular and Developmental Biology, NEI, NIH, Bldg. 6, Rm. 201, 6 Center Dr., Bethesda, MD 20892. Tel.: 301-496-9467; Fax: 301-402-0781; E-mail: joramp@nei.nih.gov.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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