Decidual/trophoblast prolactin-related protein: characterization of gene structure and cell-specific expression.
Decidual/trophoblast PRL-related protein (d/tPRP) is a member of the PRL gene family and is dually expressed in uterine and placental tissues in a highly coordinated pattern during pregnancy. In the present study, we describe the isolation and characterization of the d/tPRP gene. A lambda DASH II Wistar-Kyoto rat genomic library was screened with a labeled d/tPRP complementary DNA, resulting in the isolation of two phage clones, RGLd-41 [17.7 kilobases (kb)] and RGLd-42 (15.8 kb). RGLd-41 alone was found to contain the full-length d/tPRP gene and was used for subsequent analyses. The d/tPRP gene possesses a six-exon, five-intron organization. Relative to other highly conserved members of the PRL gene family, d/tPRP contains a single small additional exon (exon 3) situated between exons 2 and 3 of the prototypical PRL gene. The region corresponding to exon 3 of d/tPRP encodes for a unique amino acid region found in a subset of PRL family members. A reverse transcription-PCR (RT-PCR) tissue survey for d/tPRP messenger RNA revealed that d/tPRP expression was restricted to decidual and trophoblast tissues. A single transcription start site 65 bp upstream of the initiation codon was identified in decidual tissue, whereas multiple transcription start sites ranging from 61-66 bp upstream of the initiation codon were detected in placental tissue. Various tissue culture systems (primary cultures and cell lines) were evaluated for d/tPRP expression and activation of a 3.96-kb d/tPRP promoter-luciferase reporter construct. Decidual, spongiotrophoblast, and trophoblast giant cell populations expressed d/tPRP and were capable of activating the d/tPRP promoter-reporter construct, whereas other cell types were ineffective. Limited d/tPRP promoter activation was noted in uterine stromal cell lines. In summary, d/tPRP possesses a unique six-exon, five-intron gene structure and exhibits cell-specific expression that is regulated at least in part by a 3.96-kb 5'-flanking region.
Orwig
K E
KE
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City 66160, USA.
Dai
G
G
Rasmussen
C A
CA
Soares
M J
MJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003656:Decidua / Q000378:metabolism*; D005091:Exons; D005260:Female; D015698:Genomic Library; D007150:Immunohistochemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D009928:Organ Specificity; D011247:Pregnancy; D011388:Prolactin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D011921:Rats, Inbred WKY; D011994:Recombinant Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014327:Trophoblasts / Q000378:metabolism*; D014599:Uterus / Q000378:metabolism
C079559:Prl8a2 protein, rat; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011388:Prolactin