Detection of hypermutated human papillomavirus type 16 genome by Next-Generation Sequencing
Detection of hypermutated human papillomavirus type 16 genome by Next-Generation Sequencing
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通过新一代测序检测超突变的人乳头瘤病毒 16 型基因组
DOI：
10.1016/j.virol.2015.08.017
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yoshizaki Tomok
中科院分区：
文献类型：
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作者：
Wakae Kousho;Aoyama Satoru;Wang Zhe;Kitamura Kouichi;Liu Guangyan;Monjurul Ahasan Md.;Koura Miki;Imayasu Mieko;Sakamoto Naoya;Muramatsu Masamichi;Aoyama Satoru;Wang Zhe;Nakamura Mitsuhiro;Kyo Satoru;Fujiwara Hiroshi;Kyo Satoru;Kondo Satoru;Yoshizaki Tomok
人乳头瘤病毒16型（HPV 16）是宫颈癌的主要原因。我们之前证明，通过体外表达APOBEC 3，HPV 16基因组中的C至T和G至A超突变会累积。为探讨HPV 16高突变的体内特征，对3例HPV 16阳性宫颈非典型增生患者的临床标本进行差异DNA变性-PCR（3D-PCR）检测，其中2例HPV 16阳性宫颈非典型增生患者的DNA高突变。一个在E2和长对照区（LCR）都积累了超突变的样本进一步进行了下一代测序，揭示了超突变遍布LCR和所有早期基因。值得注意的是，在LCR中更频繁地观察到超突变，LCR包含病毒复制起点和早期启动子。APOBEC 3在HPV 16阳性宫颈中大量表达，表明在病毒复制和转录过程中暴露的单链DNA可能是脱氨基作用的有效靶点。这些结果进一步加强了APOBEC 3在体内引入HPV 16超突变中的作用。
Human papillomavirus type 16 (HPV16) is a major cause of cervical cancer. We previously demonstrated that C-to-T and G-to-A hypermutations accumulated in the HPV16 genome by APOBEC3 expressionin vitro. To investigatein vivocharacteristics of hypermutation, differential DNA denaturation-PCR (3D-PCR) was performed using three clinical specimens obtained from HPV16-positive cervical dysplasia, and detected hypermutation from two out of three specimens. One sample accumulating hypermutations in bothE2and the long control region (LCR) was further subjected to Next-Generation Sequencing, revealing that hypermutations spread across the LCR and all early genes. Notably, hypermutation was more frequently observed in the LCR, which contains a viral replication origin and the early promoter. APOBEC3 expressed abundantly in an HPV16-positive cervix, suggesting that single-stranded DNA exposed during viral replication and transcription may be efficient targets for deamination. The results further strengthen a role of APOBEC3 in introducing HPV16 hypermutationin vivo.