Dai G, Liu B, Szpirer C, Levan G, Kwok S C, Soares M J
Department of Physiology, University of Kansas Medical Center, Kansas City 66160, USA.
Endocrinology. 1996 Nov;137(11):5009-19. doi: 10.1210/endo.137.11.8895375.
The rat placental PRL family consists of proteins structurally related to pituitary PRL. As a consequence of attempting to characterize the gene for one of the members of the family, PRL-like protein-C (PLP-C), we identified a related gene that we have termed PLP-C variant (PLP-Cv). In this study, we present information on the PLP-Cv gene and its pattern of expression. Screening of a rat genomic library with a PLP-C cDNA resulted in the isolation of four phage clones. Nucleotide sequence analysis of the clones revealed a gene, PLP-Cv, closely related but distinct from PLP-C. The PLP-Cv gene possessed a six exon/five intron organization, unique among members of the PRL family, and was localized to chromosome 17 of the rat genome, similar to other PRL family members. A PCR strategy involving primers based on the PLP-Cv gene was used to isolate a placental PLP-Cv cDNA. PLP-Cv showed 90 and 78% sequence identity with PLP-C at nucleotide and amino acid levels, respectively. Expression of PLP-Cv was restricted to the trophoblast lineage and was coordinately activated with PLP-C beginning at day 11 of gestation and continuing until term. Primer extension analysis revealed multiple putative transcription start sites. A 2.1-kilobase pair PLP-Cv promoter-luciferase reporter construct was specifically activated in differentiating rat trophoblast cells but not in other cell types. In conclusion, we have identified a new member of the PRL family possessing considerable homology to PLP-C, a unique gene structure, and displaying a trophoblast-specific pattern of transcriptional activation.
大鼠胎盘催乳素(PRL)家族由结构上与垂体PRL相关的蛋白质组成。在试图鉴定该家族成员之一——催乳素样蛋白C(PLP-C)的基因时,我们发现了一个相关基因,我们将其命名为PLP-C变体(PLP-Cv)。在本研究中,我们提供了关于PLP-Cv基因及其表达模式的信息。用PLP-C cDNA筛选大鼠基因组文库,分离出四个噬菌体克隆。对这些克隆进行核苷酸序列分析,发现了一个与PLP-C密切相关但又不同的基因PLP-Cv。PLP-Cv基因具有六个外显子/五个内含子的结构,这在PRL家族成员中是独特的,并且与其他PRL家族成员一样,定位于大鼠基因组的17号染色体上。采用基于PLP-Cv基因的引物进行PCR策略,分离出胎盘PLP-Cv cDNA。PLP-Cv在核苷酸和氨基酸水平上与PLP-C的序列同一性分别为90%和78%。PLP-Cv的表达仅限于滋养层谱系,从妊娠第11天开始与PLP-C协同激活,并持续到足月。引物延伸分析揭示了多个推定的转录起始位点。一个2.1千碱基对的PLP-Cv启动子-荧光素酶报告构建体在分化的大鼠滋养层细胞中被特异性激活,但在其他细胞类型中未被激活。总之,我们鉴定出了PRL家族的一个新成员,它与PLP-C具有相当的同源性,具有独特的基因结构,并表现出滋养层特异性的转录激活模式。