GENE STRUCTURE AND FUNCTION IN THE RAT SEMINAL VESICLE

  • Harris, Stephen E (PI)

Project: Research project

Project Details

Description

Seminal vesicle secretory proteins (SVS) IV and V expression in the rat are
under the control of androgens. The gene for SVS IV and the genes for SVS
V have been isolated from a rat gene library. The DNA sequence has been
determined for the transcription unti and flanking sequences of the SVS IV
gene. The first objective is to derive the DNA sequence of the flanking
regions and transcription unti for the SVS V genes. The aim is to compare
5'-flanking regions for potential regulatory sites in SVS IV and SVS V
genes which may have some role in androgen action. Two major Sl nuclease sensitive sites in supercoiled plasmid pSVS 3.3
(containing flanking and transcription unit for SVS IV) have been
identified. The 5'-Sl site is at or near an inverted repeat at -113 to
-117 bp from the transcription initiation site. Along with sequencing the
SVS V genes, Sl nuclease sensitive sites will also be mapped in supercoiled
plasmids containing the SVS V genes as they are isolated, mapped, and
sequenced. DNAse I hypersensitive and Sl nuclease sensitive sites in the SVS IV and V
genes will be mapped by digestion of nuclei from normal and castrate
seminal vesicle tissue. These data will be compared to sites identified by
the Sl nuclease digestion of various supercoiled plasmids containing SVS IV
and SVS V gene fragments. A major objective will be to link the SVS IV and SVS V genes to several
eukaryotic cloning vectors with dominant selection markers (e.g., pSV-neo)
and introduce the SVS genes into recipient cell lines with well
characterized androgen receptors or into primary cultures of seminal
vesicle cells. Cell lines will be established from the transfection experiments which,
when androgen is added to the media, the transcription rate of the
introduced SVS genes will significantly increase. With these cell lines
and the production of various in vitro deletion mutants and transfection of
these mutant DNA forms of the SVS genes into cell lines, the regions in the
5'-flanking, transcription unit and/or 3'-flanking that are necessary for
androgen-dependent expression may be established. These data will broaden
our knowledge base about steroid hormone action and the control of an
apparently coordinately regulated gene set.
StatusFinished
Effective start/end date12/1/8411/30/87

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Medicine(all)

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