Differences in Stability of Viral and Viral-Cellular Fusion Transcripts in HPV-Induced Cervical Cancers

GND
1304993752
Zugehörigkeit
Department of Gynecology, Jena University Hospital
Ehrig, Franziska;
GND
136906303
ORCID
0000-0002-7012-483X
Zugehörigkeit
Department of Gynecology, Jena University Hospital
Häfner, Norman;
GND
1304994309
Zugehörigkeit
Department of Gynecology, Jena University Hospital
Driesch, Corina;
Zugehörigkeit
Department of Microbiology and Infection Control, Akershus University Hospital, 1478 Lørenskog, Norway,
Kraus Christiansen, Irene;
GND
1304901750
Zugehörigkeit
Department of Gynecology, Jena University Hospital
Beer, Katrin;
ORCID
0000-0003-2430-3500
Zugehörigkeit
Oncgnostics GmbH, 07747 Jena, Germany,
Schmitz, Martina;
GND
1111567727
ORCID
0000-0002-6702-4651
Zugehörigkeit
Department of Gynecology, Jena University Hospital
Runnebaum, Ingo B.;
GND
1111821518
ORCID
0000-0003-1235-1150
Zugehörigkeit
Department of Gynecology, Jena University Hospital, Friedrich Schiller University Jena, 07747 Jena, Germany,
Dürst, Matthias

HPV-DNA integration results in dysregulation of viral oncogene expression. Because viral-cellular fusion transcripts inherently lack the viral AU-rich elements of the 3’UTR, they are considered to be more stable than episome-derived transcripts. The aim of this study is to provide formal proof for this assumption by comparing the stability of viral early transcripts derived from episomal and integrated HPV16 DNA, respectively. Full-length cDNA of three fusion transcripts comprising viral and cellular sequences in sense orientation were amplified and cloned into the adeno-viral-vector pAd/CMV/V5-DEST. The most abundant HPV16 oncogene transcript E6*I-E7-E1vE4-E5 with and without 3’UTR, served as reference and control, respectively. Human primary keratinocytes were transduced using high titer virus stocks. qRT-PCR was performed to determine mRNA stability in relation to GAPDH in the presence of actinomycin-D. In four independent transduction experiments, all three viral-cellular fusion transcripts were significantly more stable compared to the episome-derived reference. Among the three viral-cellular fusion transcripts the most stable transcript was devoid of the instability core motif “AUUUA”. Unexpectedly, there was no significant difference in the stability between the episome-derived transcripts either with or without 3’UTR, indicating that the AU-rich elements of the 3’UTR are not contributing to RNA stability. Instead, the three “AUUUA” motifs located in the untranslated region between the viral E4 and E5 genes may be responsible for the instability. This is the first report showing that authentic viral-cellular fusion transcripts are more stable than episome-derived transcripts. The longer half-life of the fusion transcripts may result in increased levels of viral oncoproteins and thereby drive the carcinogenic process.

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