Liu D, Zong C S, Wang L H
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029-6574.
J Virol. 1993 Nov;67(11):6835-40. doi: 10.1128/JVI.67.11.6835-6840.1993.
We have shown previously that the extracellular sequences of the human insulin receptor (IR) and the insulin-like growth factor I receptor (IGFR) have an inhibitory effect on protein tyrosine kinase (PTK) activity and on the biological functions of their respective Gag-receptor fusion proteins. To study the role of IGFR carboxyl sequence in modulation of the Gag-IGFR PTK and biological activities, five mutants, CM1, CM2, CM3, CM4, and CM5, containing carboxyl deletions of 17, 27, 47, 67, and 88 amino acids (aa), respectively, were constructed from the parental virus UIGFR encoding the Gag-IGFR. Deletion of up to 27 aa had little effect on the cell-transforming and PTK activities of UIGFR. Deletions of 47 aa in CM3 abolished PTK and transforming activities. Surprisingly, a further deletion of 20 aa in CM4 beyond that in CM3 reactivated the kinase and transforming activities. CM5, containing a deletion of 20 aa beyond that in CM4, had only marginal transforming and PTK activities. We conclude that deletion of the carboxyl region of the Gag-IGFR inactivates, instead of activating as in the case with Gag-IR, its transforming activity and the amino acid sequence 1250 to 1310 is essential for PTK and transforming activities. Analysis of the ability of the full-length IGFR and its mutant receptors described above to associate with phosphatidylinositol 3 kinase indicated that the association required PTK activity and tyrosine phosphorylation of the receptors and correlated well with their transforming activities. The carboxyl 88 aa are not essential for the association.
我们之前已经表明,人胰岛素受体(IR)和胰岛素样生长因子I受体(IGFR)的细胞外序列对蛋白酪氨酸激酶(PTK)活性及其各自的Gag-受体融合蛋白的生物学功能具有抑制作用。为了研究IGFR羧基序列在调节Gag-IGFR PTK和生物学活性中的作用,从编码Gag-IGFR的亲本病毒UIGFR构建了五个突变体CM1、CM2、CM3、CM4和CM5,分别缺失17、27、47、67和88个氨基酸(aa)的羧基。缺失多达27个氨基酸对UIGFR的细胞转化和PTK活性影响很小。CM3中缺失47个氨基酸消除了PTK和转化活性。令人惊讶的是,CM4中比CM3再多缺失20个氨基酸后,激酶和转化活性重新激活。CM5比CM4再多缺失20个氨基酸,其转化和PTK活性仅处于边缘水平。我们得出结论,Gag-IGFR羧基区域的缺失会使其转化活性失活,而不是像Gag-IR那样激活,并且氨基酸序列1250至1310对于PTK和转化活性至关重要。对全长IGFR及其上述突变受体与磷脂酰肌醇3激酶结合能力的分析表明,这种结合需要PTK活性和受体的酪氨酸磷酸化,并且与它们的转化活性密切相关。羧基端的88个氨基酸对于这种结合不是必需的。