Lee H, Maihle N J
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA.
Oncogene. 1998 Jun 25;16(25):3243-52. doi: 10.1038/sj.onc.1201866.
ErbB-3 is a member of the epidermal growth factor receptor (EGFR/ErbB) family. In addition to the previously reported 6.2 kb full-length and 1.4 kb truncated c-erbB3 transcripts, we have observed a 1.7 kb c-erbB3 human transcript in Northern blots that specifically hybridizes to a probe of the extracellular domain of the receptor. Using 3'-RACE we have isolated four novel c-erbB3 cDNA clones of 1.6, 1.7, 2.1 and 2.3 kb from a human ovarian carcinoma-derived cell line. All four alternate transcripts are synthesized by readthrough of an intron and use of an alternative polyadenylation signal within this intron. Identical c-erbB3 transcripts are expressed in normal human placental tissues. Expression of these alternate transcripts is tissue-specific as indicated by Northern blot and RNase protection analyses. Fibroblasts transfected with expression vectors carrying these alternate c-erbB3 cDNA clones stably express truncated ErbB-3 products. Three of these four cDNA clones express a receptor product that is secreted. Immunoprecipitation analysis of primary cultures of human ovarian carcinomas also demonstrate the expression of a 90 kDa ErbB-3 related protein that is secreted. Furthermore, we demonstrate conservation of the exon-intron junctions between members of the erbB gene family in those regions of the gene encoding the extracellular domain. This gene structure is also conserved in the c-erbB1 homologues of Drosophila and C. elegans. Growth regulatory roles for related truncated ErbB products recently have been reported. It is, therefore, possible that the products of these four alternate c-erbB3 transcripts may also play important growth regulatory roles in normal and transformed cells.
ErbB - 3是表皮生长因子受体(EGFR/ErbB)家族的成员。除了先前报道的6.2 kb全长和1.4 kb截短的c - erbB3转录本外,我们在Northern印迹中观察到一种1.7 kb的人c - erbB3转录本,它与该受体细胞外结构域的探针特异性杂交。利用3'-RACE技术,我们从人卵巢癌衍生的细胞系中分离出四个新的1.6、1.7、2.1和2.3 kb的c - erbB3 cDNA克隆。所有这四个可变转录本都是通过内含子通读并使用该内含子内的一个可变聚腺苷酸化信号合成的。相同的c - erbB3转录本在正常人胎盘组织中表达。Northern印迹和核糖核酸酶保护分析表明,这些可变转录本的表达具有组织特异性。用携带这些可变c - erbB3 cDNA克隆的表达载体转染的成纤维细胞稳定表达截短的ErbB - 3产物。这四个cDNA克隆中的三个表达一种可分泌的受体产物。对人卵巢癌原代培养物的免疫沉淀分析也证明了一种90 kDa的分泌型ErbB - 3相关蛋白的表达。此外,我们证明了在编码细胞外结构域的基因区域中,erbB基因家族成员之间外显子 - 内含子连接的保守性。这种基因结构在果蝇和秀丽隐杆线虫的c - erbB1同源物中也保守。最近有报道称相关截短的ErbB产物具有生长调节作用。因此,这四个可变c - erbB3转录本的产物在正常细胞和转化细胞中也可能发挥重要的生长调节作用。