DeTulleo L, Kirchhausen T
Graduate Program in Virology, Harvard Medical School, 200 Longwood Ave, Boston, MA 02115, USA.
EMBO J. 1998 Aug 17;17(16):4585-93. doi: 10.1093/emboj/17.16.4585.
How important is the clathrin-dependent endocytic pathway for entry of viruses into host cells? While it is widely accepted that Semliki Forest virus (SFV), an enveloped virus, requires this pathway there are conflicting data concerning the closely related Sindbis virus, as well as varying results with picornaviruses such as human rhinovirus 14 (HRV 14) and poliovirus. We have examined the entry mode of SFV, Sindbis virus, HRV 14 and poliovirus using a method that identifies single infected cells. This assay takes advantage of the observation that the clathrin-dependent endocytic pathway is specifically and potently arrested by overexpression of dynamin mutants that prevent clathrin-coated pit budding. Using HeLa cells and conditions of low multiplicity of infection to favor use of the most avid pathway of cell entry, it was found that SFV, Sindbis virus and HRV 14 require an active clathrin-dependent endocytic pathway for successful infection. In marked contrast, infection of HeLa cells by poliovirus did not appear to require the clathrin pathway.
网格蛋白依赖的内吞途径对于病毒进入宿主细胞有多重要?虽然包膜病毒塞姆利基森林病毒(SFV)需要这一途径已被广泛接受,但关于密切相关的辛德毕斯病毒存在相互矛盾的数据,而且对于诸如人鼻病毒14型(HRV 14)和脊髓灰质炎病毒等小核糖核酸病毒的研究结果也各不相同。我们使用一种可识别单个感染细胞的方法,研究了SFV、辛德毕斯病毒、HRV 14和脊髓灰质炎病毒的进入模式。该检测利用了这样一个观察结果,即网格蛋白依赖的内吞途径会被阻止网格蛋白包被小窝出芽的发动蛋白突变体的过表达特异性且有效地阻断。使用HeLa细胞并在低感染复数条件下,以利于利用最有效的细胞进入途径,结果发现SFV、辛德毕斯病毒和HRV 14成功感染需要活跃的网格蛋白依赖的内吞途径。与之形成显著对比的是,脊髓灰质炎病毒感染HeLa细胞似乎并不需要网格蛋白途径。