人乳头瘤病毒 33 型同型变体中长控制区的转录活性。

Transcriptional activity of the long control region in human papillomavirus type 33 intratype variants.

机构信息

Department of Medical Microbiology, Faculty of Medicine, University of Debrecen, Nagyerdei krt. 98, Debrecen, H-4032, Hungary.

出版信息

Virol J. 2023 Jul 17;20(1):152. doi: 10.1186/s12985-023-02114-y.

DOI:10.1186/s12985-023-02114-y

PMID:37461035

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10353102/

Abstract

BACKGROUND

High-risk human papillomaviruses (HPVs) are responsible for the development of cervical and other anogenital cancers. Intratype sequence variants of certain high-risk HPV types (e.g. 16, 18 and 31) are thought to have different oncogenic potential, partly due to nucleotide sequence variation in the viral long control region (LCR). The LCR has an important role in the regulation of viral replication and transcription. The purpose of this study was to explore sequence variation in the LCR of HPV 33 intratype variants in Hungary and to see whether there are differences in the transcriptional activities of the variants.

METHODS

The complete HPV 33 LCR was amplified from HPV 33 positive cervical samples. After sequencing the LCR variants, multiple sequence alignment and phylogenetic analyses were carried out. Representative HPV 33 LCR sequence variants were selected for cloning and functional analysis. After transient transfection of HeLa cells, luciferase reporter assays were used to analyse the transcriptional activities of different LCR variants.

RESULTS

Altogether 10 different variants were identified by sequence analysis of the HPV 33 LCR. The results of phylogenetic analysis showed that 3 variants belonged to sublineage A1, while the other 7 variants clustered with sublineage A2. Variants belonging to sublineage A2 had significantly lower transcriptional activities than variants belonging to sublineage A1. Within sublineage A2, the two variants analysed had significantly different transcriptional activities, which was shown to be caused by the A7879G variation.

CONCLUSIONS

Nucleotide variation in the HPV 33 LCR can result in altered transcriptional activity of the intratype variants. Our results can help to understand the correlation between LCR polymorphism and the oncogenic potential of HPV 33 variants.

摘要

背景

高危型人乳头瘤病毒（HPV）是导致宫颈癌和其他肛门生殖器癌症的原因。某些高危型 HPV 类型（例如 16、18 和 31）的同型内序列变异体被认为具有不同的致癌潜能，部分原因是病毒长控制区（LCR）中的核苷酸序列变异。LCR 在病毒复制和转录的调节中起重要作用。本研究的目的是探索匈牙利 HPV 33 同型内变异体 LCR 中的序列变异，并观察变异体转录活性是否存在差异。

方法

从 HPV 33 阳性宫颈样本中扩增 HPV 33 的完整 LCR。对 LCR 变异体进行测序后，进行多序列比对和系统发育分析。选择代表性的 HPV 33 LCR 序列变异体进行克隆和功能分析。瞬时转染 HeLa 细胞后，使用荧光素酶报告基因检测分析不同 LCR 变异体的转录活性。

结果

通过 HPV 33 LCR 的序列分析共鉴定出 10 种不同的变异体。系统发育分析结果表明，3 种变异体属于亚谱系 A1，而其余 7 种变异体聚类于亚谱系 A2。属于亚谱系 A2 的变异体的转录活性明显低于属于亚谱系 A1 的变异体。在亚谱系 A2 内，分析的两种变异体具有明显不同的转录活性，这是由 A7879G 变异引起的。