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J. Biol. Chem., Vol. 277, Issue 51, 49700-49706, December 20, 2002
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From the Laboratory of Molecular and Developmental Biology, NEI,
National Institutes of Health, Bethesda, Maryland 20892
The mouse Shsp/
Orientation-dependent Influence of an Intergenic
Enhancer on the Promoter Activity of the Divergently Transcribed
Mouse Shsp/
B-crystallin and
Mkbp/HspB2 Genes*
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/B-crystallin and Mkbp/HspB2
promoter activity using dual-reporter vectors in transient transfection
and transgenic mouse experiments. Deletion of the enhancer reduced
Shsp/B-crystallin promoter activity by 30- and 93-fold
and Mkbp/HspB2 promoter activity by 6- and
10-fold in transiently transfected mouse lens -TN4 and myoblast
C2C12 cells, respectively. Surprisingly, inversion of the enhancer
reduced Shsp/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/B-crystallin promoter
preference of the intergenic enhancer in its native context. The
orientation dependence and preferential effect of the
Shsp/B-crystallin enhancer on the
Shsp/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.
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.
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