REQUIREMENT FOR VACUOLAR PROTON-ATPASE ACTIVITY DURING ENTRY OF INFLUENZA-VIRUS INTO CELLS
REQUIREMENT FOR VACUOLAR PROTON-ATPASE ACTIVITY DURING ENTRY OF INFLUENZA-VIRUS INTO CELLS
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DOI：
10.1128/jvi.69.4.2306-2312.1995
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发表时间：
1995-04-01
影响因子：
5.4
通讯作者：
CARRASCO, L
中科院分区：
文献类型：
--
作者：
GUINEA, R;CARRASCO, L
通过使用大环内酯类抗生素巴弗洛霉素A1和康卡那霉素A作为液泡质子-ATP酶（v-[H+] ATP酶）的选择性抑制剂，分析了内体酸化在流感病毒进入MDCK细胞期间所起的作用，该酶负责内体酸化。巴弗洛霉素A1和康卡那霉素A分别以5 × 10（-7）和5 × 10（-9）M的低浓度存在，当在感染的最初几分钟内加入时，可以阻止流感病毒进入细胞。病毒颗粒附着在细胞表面不是巴弗洛霉素A1作用的靶点。N，N '-二环己基碳二亚胺，质子ATP酶的非特异性抑制剂，也阻断病毒进入，而elaiophylin，血浆质子ATP酶的抑制剂，没有效果。在不同pH值的培养基中测定了巴弗洛霉素A1和康卡那霉素A的抑菌作用。当病毒在低pH条件下加入时，这两种抗生素都能有效地防止流感病毒感染。如果在加入温热的低pH培养基之前，病毒在4 ℃下与细胞结合，则这种抑制作用会降低。此外，在酸性pH下孵育细胞可以有效地阻断流感病毒感染，即使在没有抗生素的情况下。这些结果表明，pH梯度，而不是低pH，是流感病毒有效进入细胞所必需的。
The role that endosomal acidification plays during influenza virus entry into MDCK cells has been analyzed by using the macrolide antibiotics bafilomycin A1 and concanamycin A as selective inhibitors of vacuolar proton-ATPase (v-[H+]ATPase), the enzyme responsible for the acidification of endosomes. Bafilomycin A1 and concanamycin A, present at the low concentrations of 5 x 10(-7) and 5 x 10(-9) M, respectively, prevented the entry of influenza virus into cells when added during the first minutes of infection. Attachment of virion particles to the cell surface was not the target for the action of bafilomycin A1. N,N'-Dicyclohexylcarbodiimide, a nonspecific inhibitor of proton-ATPases, also blocked virus entry, whereas elaiophylin, an inhibitor of the plasma-proton ATPase, had no effect. The inhibitory actions of bafilomycin A1 and concanamycin A were tested in culture medium at different pHs. Both antibiotics powerfully prevented influenza virus infection when the virus was added under low-pH conditions. This inhibition was reduced if the virus was bound to cells at 4 degrees C prior to the addition of warm low-pH medium. Moreover, incubation of cells at acidic pH potently blocked influenza virus infection, even in the absence of antibiotics. These results indicate that a pH gradient, rather than low pH, is necessary for efficient entry of influenza virus into cells.