Koike S, Horie H, Ise I, Okitsu A, Yoshida M, Iizuka N, Takeuchi K, Takegami T, Nomoto A
Department of Microbiology, Tokyo Metropolitan Institute of Medical Science, Japan.
EMBO J. 1990 Oct;9(10):3217-24. doi: 10.1002/j.1460-2075.1990.tb07520.x.
Both genomic and complementary DNA clones encoding poliovirus receptors were isolated from genomic and complementary DNA libraries prepared from HeLa S3 cells, respectively. Nucleotide sequence analysis of these cloned DNAs revealed that the poliovirus receptor gene is approximately 20 kb long and contains seven introns in the coding region, and that at least four mRNA isoforms referring to the coding sequence are generated by alternative splicing and appear to encode four different molecules, that is, PVR alpha, PVR beta, PVR gamma and PVR delta. The predicted amino acid sequences indicate that PVR alpha and PVR delta, corresponding to the previously described cDNA clones H20A and H20B, respectively, are integral membrane proteins while the other two molecules described here for the first time lack a putative transmembrane domain. Mouse cell transformants carrying PVR alpha were permissive for poliovirus infection, but those carrying PVR beta were hardly permissive. In contrast to PVR alpha, PVR beta was not detected on the surface of the mouse cell transformants but was detected in the culture fluid by an immunological method using a monoclonal antibody against poliovirus receptor. Three types of splicing products for PVR alpha, PVR beta and PVR gamma were detected by polymerase chain reactions using appropriate primers in poly(A)+ RNAs of the brain, leukocyte, liver, lung and placenta of humans; the choice of primers used did not permit detection of PVR delta. In situ hybridization using a cDNA fragment as a probe demonstrated that the PVR gene is located at the band q13.1----13.2 of human chromosome 19.
分别从HeLa S3细胞制备的基因组文库和互补DNA文库中分离出编码脊髓灰质炎病毒受体的基因组克隆和互补DNA克隆。对这些克隆DNA进行核苷酸序列分析表明,脊髓灰质炎病毒受体基因长度约为20 kb,编码区含有7个内含子,并且通过可变剪接产生至少4种涉及编码序列的mRNA异构体,似乎编码4种不同的分子,即PVRα、PVRβ、PVRγ和PVRδ。预测的氨基酸序列表明,分别对应于先前描述的cDNA克隆H20A和H20B的PVRα和PVRδ是整合膜蛋白,而这里首次描述的其他两种分子缺乏推定的跨膜结构域。携带PVRα的小鼠细胞转化体对脊髓灰质炎病毒感染敏感,但携带PVRβ的转化体几乎不敏感。与PVRα相反,在小鼠细胞转化体表面未检测到PVRβ,但使用抗脊髓灰质炎病毒受体的单克隆抗体通过免疫方法在培养液中检测到了PVRβ。使用合适的引物通过聚合酶链反应在人脑、白细胞、肝脏、肺和胎盘的poly(A)+ RNA中检测到了PVRα、PVRβ和PVRγ的三种剪接产物;所用引物的选择不允许检测到PVRδ。使用cDNA片段作为探针的原位杂交表明,PVR基因位于人类19号染色体的q13.1----13.2带。