A regulatory element within the uteroferrin gene 5'-flanking region binds a pregnancy-associated uterine endometrial protein.
DNA-protein interactions within two putative regulatory regions distal from the transcription initiation site of the porcine uteroferrin (UF) gene were characterized. These regions, termed XB (-1,600 to -1,129 bp) and AB (-1,128 to -893 bp) exhibited transcriptional enhancer activities within the context of the heterologous SV40 promoter, that were specific to endometrial cells. DNase I and gel-shift assays demonstrated that both fragments contain a heptamer motif TGCTAGA that binds a nuclear protein present in crude and DEAE-fractionated nuclear extracts from porcine endometrium of pregnancy. This heptad sequence, designated as endometrial-associated sequence (EAS), is different from previously described nuclear protein-binding consensus sequences. Mutations in the heptamer motif abolished binding to the nuclear factor, as detected by gel-shift assays. The endometrial nuclear protein that interacts with the heptamer was characterized by Southwestern and UV cross-linking analysis. The protein has an approximate M(r) of 80 kD, is basic (pI 7.7-8.6) and is present in pig endometrium throughout pregnancy. The functional relevance of this DNA-binding protein in the control of UF gene transcription in the endometrium is discussed.
Animal Science Department, University of Florida, Gainesville 32611.
B Y
BY
Gonzalez
F J
FJ
Michel
R C
RC
Simmen
D000135:Acid Phosphatase; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004717:Endometrium / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D007527:Isoenzymes; D008667:Metalloproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000378:metabolism*; D011485:Protein Binding; D012045:Regulatory Sequences, Nucleic Acid; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D007527:Isoenzymes; D008667:Metalloproteins; D009687:Nuclear Proteins; D011257:Pregnancy Proteins; D004247:DNA; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase