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从足底疣和感染的上皮细胞培养物中回收的人乳头瘤病毒1型RNA转录本的鉴定与定位

Identification and mapping of human papillomavirus type 1 RNA transcripts recovered from plantar warts and infected epithelial cell cultures.

作者信息

Chow L T, Reilly S S, Broker T R, Taichman L B

出版信息

J Virol. 1987 Jun;61(6):1913-8. doi: 10.1128/JVI.61.6.1913-1918.1987.

Abstract

Multiple spliced transcripts of human papillomavirus type 1 were detected by electron microscopic analysis of R-loops formed with total RNA extracted from plantar warts and with poly(A)+ RNA isolated from cultured keratinocytes infected with human papillomavirus type 1. The 5' ends of the RNAs were mapped to sites in the E7 open reading frame (ORF), just upstream of the E6 ORF and in the upstream regulatory region. Species with 5' ends in E7 accounted for over 95% of all transcripts seen. Two polyadenylation sites were used, one at the end of the early (E) region of the viral DNA, the other at the end of the late (L) region. The most abundant species had a short 5' exon of approximately 100 nucleotides spanning the junction of the E7 and E1 ORFs spliced to a 3' exon of 800 nucleotides in the region with overlapping E2 and E4 ORFs; it was polyadenylated at the end of the E region. This species probably encodes the abundant E4 protein found in plantar warts (F. Breitburd, O. Croissant, and G. Orth, Cancer Cells, vol. 5, in press; J. Doorbar, D. Campbell, R. J. A. Grand, and P. H. Gallimore, EMBO J. 5:355-362, 1986). Other transcripts had exons spanning the E6-E7 ORFs, the E4-E5-L2-L1 ORFs, or the L1 ORF. The infrequent L1 transcript, probably the mRNA coding for the major capsid protein, had the same 5' exon in E7 as the abundant mRNA spliced from E1 and E4 ORFs, suggesting genetic regulation via the choice of the alternative polyadenylation sites or mRNA processing.

摘要

通过对从足底疣中提取的总RNA以及从感染了1型人乳头瘤病毒的培养角质形成细胞中分离出的聚腺苷酸加尾RNA(poly(A)+ RNA)所形成的R环进行电子显微镜分析,检测到了1型人乳头瘤病毒的多个剪接转录本。RNA的5'端定位于E7开放阅读框(ORF)中的位点,就在E6 ORF上游以及上游调控区域。5'端位于E7的转录本种类占所有可见转录本的95%以上。使用了两个聚腺苷酸化位点,一个在病毒DNA早期(E)区域末端,另一个在晚期(L)区域末端。最丰富的转录本种类有一个约100个核苷酸的短5'外显子,跨越E7和E1 ORF的交界处,剪接到E2和E4 ORF重叠区域中一个800个核苷酸的3'外显子;它在E区域末端进行聚腺苷酸化。这种转录本可能编码在足底疣中发现的丰富的E4蛋白(F. Breitburd、O. Croissant和G. Orth,《癌细胞》,第5卷,即将发表;J. Doorbar、D. Campbell、R. J. A. Grand和P. H. Gallimore,《欧洲分子生物学组织杂志》5:355 - 362,1986年)。其他转录本的外显子跨越E6 - E7 ORF、E4 - E5 - L2 - L1 ORF或L1 ORF。罕见的L1转录本,可能是编码主要衣壳蛋白的mRNA,其在E7中的5'外显子与从E1和E4 ORF剪接而来的丰富mRNA相同,这表明通过选择替代聚腺苷酸化位点或mRNA加工进行基因调控。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8279/254198/81f266978c7c/jvirol00097-0159-a.jpg

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