Isolation and characterization of a rat epidermal growth factor related protein

摘要

A cDNA fragment clone of 1583 base pairs with 90-95% sequence homology to the mouse EGFR and a truncated rat EGFR was isolated. The isolated cDNA lacked the termination codon, rapid amplification of cDNA ends (RACE) was applied to obtain the 3-terminal end. The full-length cDNA revealed 1753 base pairs that contained 227 base pairs of 5 untranslated region and an open reading frame encoding 473 amino acids which showed an 84% and 91% homology, respectively, to a rat truncated EGFR and the mouse EGFR. We refer to the product of the newly isolated cDNA as ERRP (EGF-Receptor Related Protein). In Northern-blot analysis with poly A RNA from different rat tissues, ERRP cDNA hybridized strongly to a mRNA transcript of about 1.8 kb. Maximal expression was noted in the small intestine, followed by colon, liver, gastric mucosa and other tissues. Transfection of ERRP cDNA in HCT-116 cells, a colon cancer cell line, markedly inhibits proliferation in monolayer and in soft agar and attenuates EGF-R tyrosine kinase activity. Transfection of ERRP in HCT-116 also delays tumor growth in SCID mice injected with these cells compared to the vector-transfected control cells. On the other hand, proliferation of the vector-transfected control, but not ERRP transfected HCT cells could be stimulated by TGF- (10¹⁰M and 10⁹M). The ERRP cDNA represents a new member of the EGFR gene family, and its protein product plays a key role in modulating the function of EGFR.

机译：分离出1583个碱基对的cDNA片段克隆，与小鼠EGFR具有90-95％的序列同源性，并分离了截短的大鼠EGFR。分离的cDNA缺少终止密码子，应用cDNA末端的快速扩增（RACE）获得3-末端。全长cDNA揭示了1753个碱基对，其中包含227个碱基对的5个非翻译区和一个开放阅读框，该可读框编码473个氨基酸，与大鼠截短的EGFR和小鼠EGFR的同源性分别为84％和91％。我们将新分离的cDNA的产物称为ERRP（EGF受体相关蛋白）。在用来自不同大鼠组织的poly A RNA进行Northern印迹分析时，ERRP cDNA与约1.8 kb的mRNA转录物强烈杂交。在小肠中发现最大的表达，其次是结肠，肝，胃粘膜和其他组织。 ERRP cDNA在结肠癌细胞系HCT-116细胞中的转染显着抑制单层和软琼脂中的增殖，并减弱EGF-R酪氨酸激酶活性。与载体转染的对照细胞相比，HCT-116中ERRP的转染也延迟了注射了这些细胞的SCID小鼠的肿瘤生长。另一方面，TGF-（10 ^{10 M和10 ^{9 M）可以刺激载体转染的对照细胞的增殖，而不是ERRP转染的HCT细胞的增殖。 ERRP cDNA代表EGFR基因家族的新成员，其蛋白质产物在调节EGFR功能中起关键作用。}}

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公开/公告号US2002045215A1

专利类型
公开/公告日2002-04-18

原文格式PDF
申请/专利权人 MAJUMDAR ADHIP P.N.;
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申请/专利号US20010867521
发明设计人 ADHIP P.N. MAJUMDAR;
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申请日2001-05-31
分类号C12P21/04;C12N15/85;C07H21/04;C12N5/08;
国家 US
入库时间 2022-08-22 00:52:32

Isolation and characterization of a rat epidermal growth factor related protein

摘要

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