Papillomavirus Infection Requires γ Secretase
Papillomavirus Infection Requires γ Secretase
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DOI：
10.1128/jvi.01081-10
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发表时间：
2010-10-01
影响因子：
5.4
通讯作者：
Roden, Richard B. S.
中科院分区：
文献类型：
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作者：
Karanam, Balasubramanyam;Peng, Shiwen;Roden, Richard B. S.
乳头状瘤病毒突破细胞膜将其基因组货物运送到细胞核的机制知之甚少。在这里，我们表明，感染范围广泛的乳头瘤病毒类型需要膜内蛋白酶γ分泌酶。γ-分泌酶抑制剂（S，S）-2-[2-（3，5-二氟苯基）-乙酰氨基]- N-（1-甲基-2-氧代-5-苯基-2，3-二氢-1H-苯并[e][1，4]二氮杂卓-3-基）-丙酰胺（化合物XXI）在体外抑制测试的所有类型的乳头瘤病毒假病毒体的感染，50%抑制浓度（IC 50）为130至1，000 pM，无论报告基因构建体如何并且不影响细胞活力。相反，XXI不抑制由BK或默克尔细胞多瘤病毒衍生的腺病毒或假病毒体的体外感染。阴道应用XXI可预防人乳头瘤病毒16型（HPV 16）假病毒体感染小鼠生殖道。Nicastrin和早老素-1是γ-分泌酶复合物的重要组成部分，缺乏这些组成部分中的任何一种的小鼠胚胎成纤维细胞都不会被HPV 16感染，而野生型和β-分泌酶（BACE 1）缺陷细胞则易感。HPV 16进入Lamp-1阳性核周囊泡的摄取和衣壳的解体都不依赖于γ-分泌酶活性，而衣壳解体则揭示了内部L1和L2表位以及溴脱氧尿苷（BrdU）标记的双核苷酸DNA。然而，XXI对γ-分泌酶活性的阻断阻止了HPV 16标记的BrdU标记的DNA到达ND 10亚核结构域。由于先前的研究表明，L2是至关重要的内体逃逸和靶向ND 10的病毒DNA和γ分泌酶位于内体膜，我们的研究结果表明，无论是L2或细胞内受体裂解的γ分泌酶作为乳头瘤病毒逃逸内体。
The mechanism by which papillomaviruses breach cellular membranes to deliver their genomic cargo to the nucleus is poorly understood. Here, we show that infection by a broad range of papillomavirus types requires the intramembrane protease gamma secretase. The gamma-secretase inhibitor (S, S)-2-[2-(3,5-difluorophenyl)-acetylamino]- N-(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4] diazepin-3-yl)-propionamide (compound XXI) inhibits infection in vitro by all types of papillomavirus pseudovirions tested, with a 50% inhibitory concentration (IC50) of 130 to 1,000 pM, regardless of reporter construct and without impacting cellular viability. Conversely, XXI does not inhibit in vitro infection by adenovirus or pseudovirions derived from the BK or Merkel cell polyomaviruses. Vaginal application of XXI prevents infection of the mouse genital tract by human papillomavirus type 16 (HPV16) pseudovirions. Nicastrin and presenilin-1 are essential components of the gamma-secretase complex, and mouse embryo fibroblasts deficient in any one of these components were not infected by HPV16, whereas wild-type and beta-secretase (BACE1)-deficient cells were susceptible. Neither the uptake of HPV16 into Lamp-1-positive perinuclear vesicles nor the disassembly of capsid to reveal both internal L1 and L2 epitopes and bromodeoxyuridine (BrdU)-labeled encapsidated DNA is dependent upon gamma-secretase activity. However, blockade of gamma-secretase activity by XXI prevents the BrdU-labeled DNA encapsidated by HPV16 from reaching the ND10 subnuclear domains. Since prior studies indicate that L2 is critical for endosomal escape and targeting of the viral DNA to ND10 and that gamma secretase is located in endosomal membranes, our findings suggest that either L2 or an intracellular receptor are cleaved by gamma secretase as papillomavirus escapes the endosome.