Characterization of the gene encoding a folate-binding protein expressed in human placenta. Identification of promoter activity in a G-rich SP1 site linked with the tandemly repeated GGAAG motif for the ets encoded GA-binding protein.
The gene encoding a folate-binding protein (FBP) expressed in human placenta has been cloned by screening a genomic library with the KB cell FBP complementary DNA. This gene, contained in a 10-kilobase EcoRI fragment of this genomic clone, has 5 exons, 4 introns, the AATAA polyadenylation signal in the 3'-untranslated region, and a 5'-flanking sequence which contains the promoter elements, all of which span approximately 5 kilobases. Transcription initiation was mapped by RNase protection to a site 73 base pairs downstream from a G-rich sequence linked to a tandemly repeated GGAAG sequence which is a motif that the ets oncogene encoded GA-binding protein (GABP) transcription factor binds. Gel-shift and supershift mobility assays indicate that the G-rich sequence and the ets motif bind specifically to SP1 and GABP, respectively. These cis regulatory elements in tandem drive expression of the chloramphenicol acetyltransferase reporter gene in transiently transfected mouse 3T3 cells. The location of these elements upstream of transcription initiation in this gene, which lacks an appropriately located TATA box promoter, indicates that this SP1-GA binding region most probably regulates expression of this placental FBP. The gene encoding this placental FBP has been assigned the FBP/PL-1 gene because it is a member of a multigene family that includes a gene encoding a FBP expressed in both KB cells and placenta and its unprocessed pseudogene.
Sadasivan
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E
Department of Medicine, State University of New York Health Science Center, Brooklyn.
Cedeno
M M
MM
Rothenberg
S P
SP
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism*; D005091:Exons; D005260:Female; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism; D051268:GA-Binding Protein Transcription Factor; D006801:Humans; D007438:Introns; D007624:KB Cells; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D009857:Oncogenes; D010920:Placenta / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D012326:RNA Splicing; D011956:Receptors, Cell Surface; D015682:Retroviridae Proteins, Oncogenic / Q000096:biosynthesis / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D013379:Substrate Specificity; D014157:Transcription Factors / Q000096:biosynthesis / Q000378:metabolism*
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D058973:Folate Receptors, GPI-Anchored; D051268:GA-Binding Protein Transcription Factor; D009838:Oligodeoxyribonucleotides; D011956:Receptors, Cell Surface; D015682:Retroviridae Proteins, Oncogenic; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; C050379:oncogene proteins v-ets; D004247:DNA; D005492:Folic Acid