# ESTROGEN REGULATION OF THE RAT LH BETA GENE

> **NIH NIH R01** · UNIVERSITY OF VIRGINIA · 1993 · $145,404

## Abstract

Lutropin (LH), a pituitary glycoprotein hormone, plays a critical
role in gonadal development and function, and regulates the
production of sex steroids such as estradiol (E2).  LH consists of
two subunits, the alpha subunit common to all glycoprotein
hormones, and the unique LHBeta subunit which is under more
stringent physiological regulation.  LH production in vivo is under
both positive and negative regulation by E2, but it is unclear
whether this feedback occurs at the hypothalamic or pituitary
level,  or both.  We have shown that the transcription rate of the
kLHBeta gene is negatively regulated by E2 in vivo, but preliminary
experiments indicate E2 stimulation of transcription in pituitary
fragments and in transient expression assays.  The first aim is to
define the direct actions of E2 on the LHBeta gene by measuring E2-
regulated transcription free of hypothalamic influences.  The time
and dose dependence of the E2 response will be assessed in isolated
pituitary fragments and in cell culture in the absence and presence
of gonadotropin releasing hormone (GnRH).  The second aim is to
identify regions of the LHBeta gene which confer an E2 response.
Filter-binding assays with labeled gene fragments and purified E2
receptor complex indicate at least one putative receptor binding
site in the 5'-flanking region of the LHBeta gene.  This region and
other potential receptor binding sites will be studied by
Exonuclease III and DNAse I footprinting studies, as well as by
mutational analysis, to identify the sequence for receptor binding.
Gene regions necessary for the biological E2 response will be
identified by transient expression assays, in which upstream LHBeta
gene regions inserted next to a thymidine kinase or prolactin
promoter controlling expression of a reporter gene are transfected
into GH3 cells or normal pituitary cells.  It is anticipated that
the receptor binding region will be required for the response, but
other DNA elements and binding proteins may be necessary, and will
be identified.  The ability of specific gene regions to confer the
E2 response to the reporter gene, and the sequence and positioning
of these elements will be determined by deletion analysis.  The
importance of specific nucleotides within these sequences will be
evaluated by site-directed mutagenesis.  The ability of other
hormones to modulate the E2 response will be tested.  The
interaction of the antiestrogen tamoxifen with LHBeta gene E2
regulatory regions will be measured by tamoxifen-E2 receptor
footprint analysis, and tamoxifen and progesterone modulation of
the E2 effects will be investigated by transcription, and in
transient expression assays.  Reporter gene constructs containing
the homologous LHBeta gene promoter and 5'flanking regions will be
transfected into pituitary cells and tested for expression and E2
regulation in the absence or presence of GnRH.  These studies will
add to our understanding of the direct E2 effects on  LH production
and clarify ...

## Key facts

- **NIH application ID:** 2199687
- **Project number:** 5R01HD025719-05
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **SAM.gov UEI:** JJG6HU8PA4S5
- **Principal Investigator:** MARGARET A SHUPNIK
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $145,404
- **Award type:** 5
- **Project period:** 1989-06-01 → 1995-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2199687

## Citation

> US National Institutes of Health, RePORTER application 2199687, ESTROGEN REGULATION OF THE RAT LH BETA GENE (5R01HD025719-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2199687. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*