Zhang Q K, Boast S, de los Santos K, Begemann M, Goff S P
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
J Virol. 1996 Nov;70(11):8089-97. doi: 10.1128/JVI.70.11.8089-8097.1996.
Retroviral genomes encoding a portion of the Moloney murine leukemia virus Gag protein fused to portions of the murine axl cDNA were constructed so as to mimic naturally occurring transforming viruses. Virus MA1 retained 5 amino acids of the extracellular domain and the complete transmembrane and intracellular domains of Axl; virus MA2 retained only the intracellular Axl sequences beginning 33 amino acids downstream of the transmembrane region. Although both viruses could transform NIH 3T3 cells, they induced different morphological changes. MA1 transformants became elongated and assumed a cross-hatched pattern, while MA2 transformants were round and very refractile and grew to high density. Gag-Axl and Glyco-Gag-Axl proteins were detected in both types of transformed cells and were predominantly localized to the cytoplasmic compartment. When cell-free v-axl virus supernatants were introduced into wild-type BALB/c neonates, Rag-2-deficient mice, or c-myc transgenic mice, they did not cause tumors in a 3-month period. However, MA2-transformed NIH 3T3 cells, but not MA1 or control cells, could establish sarcomas by subcutaneous or intraperitoneal injection into BALB/c neonates. These results show that the transforming potential of the axl gene can be activated by truncation of the extracellular domain of the receptor and fusion of the remaining sequence to the gag gene.
构建了编码与鼠axl cDNA部分融合的莫洛尼鼠白血病病毒Gag蛋白部分的逆转录病毒基因组,以模拟天然存在的转化病毒。病毒MA1保留了Axl细胞外结构域的5个氨基酸以及完整的跨膜和细胞内结构域;病毒MA2仅保留了从跨膜区域下游33个氨基酸开始的细胞内Axl序列。尽管两种病毒都能转化NIH 3T3细胞,但它们诱导了不同的形态变化。MA1转化细胞变长并呈现交叉阴影模式,而MA2转化细胞呈圆形且非常有折光性,并生长至高密度。在两种类型的转化细胞中均检测到Gag-Axl和糖基化Gag-Axl蛋白,且主要定位于细胞质区室。当将无细胞的v-axl病毒上清液引入野生型BALB/c新生小鼠、Rag-2缺陷小鼠或c-myc转基因小鼠时,在3个月内它们未引发肿瘤。然而,MA2转化的NIH 3T3细胞(而非MA1或对照细胞)通过皮下或腹腔注射到BALB/c新生小鼠中可形成肉瘤。这些结果表明,axl基因的转化潜能可通过受体细胞外结构域的截短以及剩余序列与gag基因的融合而被激活。