Analysis of transcription and estrogen insensitivity in the female mouse after targeted disruption of the estrogen receptor gene
- PMID: 8584021
- DOI: 10.1210/mend.9.11.8584021
Analysis of transcription and estrogen insensitivity in the female mouse after targeted disruption of the estrogen receptor gene
Abstract
We employed homologous recombination in mouse embryonic stem cells to disrupt the estrogen receptor (ER) gene. Subsequently generated mice that are homozygous for the gene disruption, termed ERKO, possess no demonstrable wild-type ER by Western blot analysis. However, the presence of residual high affinity binding, as detected by [3H]estradiol binding assays and sucrose gradients in uterine extracts from ERKO females prompted further investigation of transcription and translation products from the disrupted ER gene. Analysis of ERKO uterine messenger RNA (mRNA) by reverse transcriptase-polymerase chain reaction demonstrated that although no full-length wild-type ER mRNA was present, two smaller transcripts, labeled E1 and E2, were identified and partially sequenced. Both ERKO transcripts are splicing variants that result in the disrupting NEO sequence being partially or completely removed from the mRNA. In the ERKO-E2 variant, this results in a frame shift and the creation of at least two stop codons downstream. In the ERKO-E1 variant, the ER reading frame is preserved and encodes for a smaller mutant ER that could be the source of the residual estradiol binding. When this mutant form is overexpressed and characterized in vitro, it results in a smaller protein of the predicted size that possesses significantly reduced estrogen-dependent transcriptional activity compared with that of the wild-type ER. Despite residual amounts of an impaired ER variant, estrogen insensitivity in the female ERKOs was confirmed by the failure of estrogen treatment to induce known uterine markers of estrogen action, such as increased DNA synthesis, and transcription of the progesterone receptor, lactoferrin, and glucose-6-phosphate dehydrogenase genes. Furthermore, serum levels of estradiol in the ERKO female are more than 10-fold higher than those in the wild type, consistent with a syndrome of hormone insensitivity.
Similar articles
-
Estrogen receptor gene disruption: molecular characterization and experimental and clinical phenotypes.Recent Prog Horm Res. 1996;51:159-86; discussion 186-8. Recent Prog Horm Res. 1996. PMID: 8701078 Review.
-
Methoxychlor stimulates estrogen-responsive messenger ribonucleic acids in mouse uterus through a non-estrogen receptor (non-ER) alpha and non-ER beta mechanism.Endocrinology. 1999 Aug;140(8):3526-33. doi: 10.1210/endo.140.8.6877. Endocrinology. 1999. PMID: 10433208
-
Disruption of estrogen signaling does not prevent progesterone action in the estrogen receptor alpha knockout mouse uterus.Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3646-51. doi: 10.1073/pnas.96.7.3646. Proc Natl Acad Sci U S A. 1999. PMID: 10097091 Free PMC article.
-
Function of the exon 7 deletion variant estrogen receptor alpha protein in an estradiol-resistant, tamoxifen-sensitive human endometrial adenocarcinoma grown in nude mice.Gynecol Oncol. 2002 Feb;84(2):271-9. doi: 10.1006/gyno.2001.6509. Gynecol Oncol. 2002. PMID: 11812086
-
Estrogen receptor null mice: what have we learned and where will they lead us?Endocr Rev. 1999 Jun;20(3):358-417. doi: 10.1210/edrv.20.3.0370. Endocr Rev. 1999. PMID: 10368776 Review.
Cited by 126 articles
-
About three-fourths of mouse proteins unexpectedly appear at a low position of SDS-PAGE, often as additional isoforms, questioning whether all protein isoforms have been eliminated in gene-knockout cells or organisms.Protein Sci. 2020 Apr;29(4):978-990. doi: 10.1002/pro.3823. Epub 2020 Jan 23. Protein Sci. 2020. PMID: 31930537
-
Uterine glycolytic enzyme expression is affected by knockout of different estrogen receptor subtypes.Biomed Rep. 2019 Oct;11(4):135-144. doi: 10.3892/br.2019.1234. Epub 2019 Aug 20. Biomed Rep. 2019. PMID: 31565219 Free PMC article.
-
Disruptions in the female reproductive system on consumption of calcium carbide ripened fruit in mouse models.Heliyon. 2019 Sep 6;5(9):e02397. doi: 10.1016/j.heliyon.2019.e02397. eCollection 2019 Sep. Heliyon. 2019. PMID: 31517122 Free PMC article.
-
Progesterone and Estrogen Signaling in the Endometrium: What Goes Wrong in Endometriosis?Int J Mol Sci. 2019 Aug 5;20(15):3822. doi: 10.3390/ijms20153822. Int J Mol Sci. 2019. PMID: 31387263 Free PMC article. Review.
-
A critical role for estrogen signaling in penis development.FASEB J. 2019 Sep;33(9):10383-10392. doi: 10.1096/fj.201802586RR. Epub 2019 Jun 21. FASEB J. 2019. PMID: 31225966 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
-
Full Text Sources
-
Other Literature Sources
-
Medical
-
Molecular Biology Databases
-
Research Materials