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J. Biol. Chem., Vol. 277, Issue 50, 48771-48778, December 13, 2002
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From the Department of Immunology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030
The epididymis is a useful model system to
understand the mechanisms that govern region-specific gene expression,
as many gene products display spatially restricted expression within
this organ. However, surprisingly little is known about how this
regulation is achieved. Here, we report regulatory sequences from the
Pem homeobox gene that drive expression in different
subregions of the mouse epididymis in vivo. We found that
the 0.3-kb 5'-flanking sequence (region I) from the Pem
proximal promoter (Pem Pp) was sufficient to confer
androgen-dependent and developmentally regulated expression
in the caput region of the epididymis. Expression was restricted to the
normal regions of expression of Pem in the caput (segments 2-4), but
there was also aberrant expression in the corpus region. This corpus
misexpression was extinguished when 0.6 kb of Pem Pp
5'-flanking sequence was included in the transgene, indicating that one
or more negative regulatory elements exist between 0.6 and 0.3 kb
upstream of the Pem Pp start site (region II). When
heterologous sequences were introduced upstream of the Pem
Pp, expression was further restricted, mainly to caput segment 3, implying that the Pem Pp has segment-specific regulatory
elements. To our knowledge, the regulatory regions we have identified
are the shortest so far defined that dictate regionally localized expression in the epididymis in vivo. They may be useful
for identifying the factors that regulate region-specific expression in
the epididymis, for expressing and conditionally knocking out genes in
different subregions of the epididymis, for treating male infertility,
and for generating novel methods of male contraception.
Pem Homeobox Gene Regulatory Sequences That
Direct Androgen-dependent Developmentally Regulated
Gene Expression in Different Subregions of the Epididymis*
*
This work was supported by National Institutes of Health
Grant CA78023.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: Dept. of Immunology,
Box 180, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030. Tel.: 713-794-5526; Fax: 713-745-0846; E-mail: mwilkins@mdanderson.org.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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