VITRONECTIN RECEPTOR ANTIBODIES INHIBIT INFECTION OF HELA AND A549 CELLS BY ADENOVIRUS-TYPE-12 BUT NOT BY ADENOVIRUS TYPE-2
VITRONECTIN RECEPTOR ANTIBODIES INHIBIT INFECTION OF HELA AND A549 CELLS BY ADENOVIRUS-TYPE-12 BUT NOT BY ADENOVIRUS TYPE-2
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DOI：
10.1128/jvi.68.9.5925-5932.1994
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发表时间：
1994-09-01
影响因子：
5.4
通讯作者：
FREIMUTH, P
中科院分区：
文献类型：
--
作者：
BAI, M;CAMPISI, L;FREIMUTH, P
12 型腺病毒 (Ad12) 的五邻体碱基基因已被测序并编码 497 个残基的多肽，比 Ad2 的五邻体碱基短 74 个残基。 Ad2 和 Ad12 蛋白在氨基末端和羧基末端高度保守，但在中心区域截然不同，其中 Ad12 序列缺失 63 个残基。该可变区内保守的是 Arg-Gly-Asp (RGD) 序列，该序列在 Ad2 五邻体碱基中与靶细胞膜中的整合素结合，从而提高感染的速率或效率。 Ad12五邻体碱基在大肠杆菌中表达，纯化的重折叠蛋白在体外与Ad2纤维组装。与 Ad2 五邻体碱基相反，Ad12 蛋白未能导致贴壁细胞变圆或促进 HeLa S3 悬浮细胞贴壁；然而，A549 细胞确实附着在涂有任一蛋白质的表面上，并且用整合素 alpha v beta 3 单克隆抗体对细胞进行预处理，从而降低了与背景水平的附着。用整联蛋白αvβ3或αvβ5单克隆抗体或用Ad2五邻体碱基蛋白的含有RGD的片段处理HeLa ind A549细胞抑制了Ad12的感染，但对Ad2的感染没有影响，并且在某些情况下增强了Ad2的感染。纯化的 Ad2 纤维蛋白将放射性标记的 Ad2 和 Ad12 病毒体与 HeLa 和 A549 细胞的结合降低到接近背景水平，但强烈抑制 Ad2 感染的纤维浓度仅微弱地抑制 Ad12 感染。这些数据表明，单独含有αv的整联蛋白可能足以支持Ad12的感染，并且Ad2不能有效地利用该途径。
The penton base gene from adenovirus type 12 (Ad12) was sequenced and encodes a 497-residue polypeptide, 74 residues shorter than the penton base from Ad2. The Ad2 and Ad12 proteins are highly conserved at the amino- and carboxy-terminal ends but diverge radically in the central region, where 63 residues are missing from the Ad12 sequence. Conserved within this variable region is the sequence Arg-Gly-Asp (RGD), which, in the Ad2 penton base, binds to integrins in the target cell membrane, enhancing the rate or the efficiency of infection. The Ad12 penton base was expressed in Escherichia coli, and the purified refolded protein assembled in vitro with Ad2 fibers. In contrast to the Ad2 penton base, the Ad12 protein failed to cause the rounding of adherent cells or to promote attachment of HeLa S3 suspension cells; however, A549 cells did attach to surfaces coated vith either protein and pretreatment of the cells with an integrin alpha v beta 3 monoclonal antibody reduced attachment to background levels. Treatment of HeLa ind A549 cells with integrin alpha v beta 3 or alpha v beta 5 monoclonal antibodies or with an RGD-containing fragment of the Ad2 penton base protein inhibited infection by Ad12 but had no effect on and in some cases enhanced infection by Ad2. Purified Ad2 fiber protein reduced the binding of radiolabeled Ad2 and Ad12 virions to HeLa and A549 cells nearly to background levels, but the concentrations of fiber that strongly inhibited infection by Ad2 only weakly inhibited Ad12 infection. These data suggest that alpha v-containing integrins alone may be sufficient to support infection by Ad12 and that this pathway is not efficiently used by Ad2.