Differential molecular mechanism of the estrogen action that regulates lactoferrin gene in human and mouse

Christina T. Teng, Youhua Liu, Nengyu Yang, David Walmer, Timothy Panella

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

The 5′-flanking region of the human lactoferrin gene was isolated from a human placental genomic library. This genomic clone contains a 16-kilobase pair (kbp) insert and produces seven fragments when digested with the SacI restriction enzyme. We sequenced one of the fragments that comprises 1294 bp of the 5′-flanking sequence, 79 bp of the first exon, and 690 bp of the first intron. A major transcription start site was mapped by primer extension. The region immediately upstream from the transcription initiation site following the first exon is abundant in G and C nucleotides. In the promoter and 5′-flanking region within a 300-bp stretch (-465 to -165) of the DNA, we found a noncanonical TATA box (ATAAA), CAAT-like sequence (CAAC) and sequences homologous to the consensus SP1 binding site, Pu.1/Sp.1 binding element (PU box), two half-palindromic estrogen response elements (EREs; GGTCA), an imperfect ERE (GGTCAAGGCGATC), and a sequence resembling the chicken ovalbumin upstream promoter transcription factor (COUP-TF) binding site (GTCTCACAGGTCA). The COUP-TF binding site and the imperfect ERE shared five nucleotides (GGTCA). With the exception of the two half-palindromic EREs, the elements with very well matched sequences were also found in the corresponding positions in the mouse lactoferrin gene. The synthetic oligonucleotide, including the 26 bp of COUP/ERE sequence, was cloned before the SV40 promoter in a chloramphenicol acetyltransferase reporter construct. These chimeric plasmids were transiently transfected into human endometrium carcinoma RL95-2 cells to assess hormone responsiveness. We found that the COUP/ERE element acted as an enhancer in response to estrogen stimulation. In vitro DNase I footprinting analysis showed binding of the estrogen receptor on the imperfect ERE. In contrast to the mouse lactoferrin COUP/ERE element, COUP-TF does not interact with this element, as demonstrated by band shift assay and site-directed mutagenesis. Therefore, the molecular mechanisms of the estrogen action that govern the lactoferrin gene expression differ between mouse and human.

Original languageEnglish (US)
Pages (from-to)1969-1981
Number of pages13
JournalMolecular Endocrinology
Volume6
Issue number11
DOIs
StatePublished - Nov 1 1992
Externally publishedYes

Fingerprint

COUP Transcription Factors
Lactoferrin
5' Flanking Region
Estrogens
Transcription Initiation Site
Binding Sites
Genes
Exons
Nucleotides
TATA Box
Chloramphenicol O-Acetyltransferase
Genomic Library
Deoxyribonuclease I
Response Elements
Endometrial Neoplasms
Sequence Homology
Site-Directed Mutagenesis
Oligonucleotides
Estrogen Receptors
Introns

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Differential molecular mechanism of the estrogen action that regulates lactoferrin gene in human and mouse. / Teng, Christina T.; Liu, Youhua; Yang, Nengyu; Walmer, David; Panella, Timothy.

In: Molecular Endocrinology, Vol. 6, No. 11, 01.11.1992, p. 1969-1981.

Research output: Contribution to journalArticle

Teng, Christina T. ; Liu, Youhua ; Yang, Nengyu ; Walmer, David ; Panella, Timothy. / Differential molecular mechanism of the estrogen action that regulates lactoferrin gene in human and mouse. In: Molecular Endocrinology. 1992 ; Vol. 6, No. 11. pp. 1969-1981.
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