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. 2022 Feb 21;58(2):98–112. doi: 10.1007/s11262-022-01889-6

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — A Linearized HPV16 genome (numbers refer to the HPV16 reference strain GeneBank: K02718.1). Early and late genes are indicated. P97: HPV16 early promoter. P670: HPV16 late promoter. Black oval: splice donor. White oval: splice acceptor. pAE: HPV16 early polyadenylation site. pAL: HPV16 late polyadenylation site. LCR: HPV16 long control region. B HPV16 subgenomic plasmid pC97ELsLuc encodes all HPV16 genes. HPV16 early promoter P97 was replaced by human cytomegalovirus immediate early promoter (CMV). Secreted luciferase (sLuc) gene was integrated in the L1 gene following the poliovirus 2A internal ribosomal entry site (IRES) sequence. C Schematic structures of HPV16 early transcripts produced from pC97ELsLuc. Splicing at SD226 occurs independently of splicing at the downstream SD880 and generates splice variants in the E6- and E7-coding region. Arrows indicate HPV16 RT-PCR primers. D Effect of ALKBH5 on HPV16 E6/E7 mRNA splicing was monitored by RT-PCR with indicated primer pair on RNA extracted from triplicate transfections of pC97ELsLuc with empty pUC plasmid (−) or pALKBH5. E Densitometric quantitation of the results in E. The percentage of each indicated cDNA isoform of all cDNAs in a lane in the absence of ALKBH5 (−) or in the presence of ALKBH5 overexpression is displayed. F Effect of FTO or ALKBH5 on HPV16 E4 mRNA splicing was monitored by RT-PCR with indicated primer pair on RNA extracted from HeLa cells transfected with pC97ELsLuc and empty pUC plasmid (−), pFTO, or pALKBH5