Characterization of cDNAs of spliced HPV-11 E2 mRNA and other HPV mRNAs recovered via retrovirus-mediated gene transfer

Mitch 0. Rotenberg, Cheng Ming Chiang, Maureen L. Ho, Thomas R. Broker, Louise T. Chows

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Human papillomaviruses (HPVs) are associated with hyperproliferations of cutaneous or mucosal epithelium. These viruses cannot be propagated in any cell culture system. Because cloning cDNA copies of HPV mRNAs recovered from human lesions has met with only very limited success, the characterization of HPV mRNAs has been problematic. Using the Moloney murine leukemia virus vector system (C. L. Cepko, B. E. Roberts, and R. C. Mulligan, 1984, Cell 37, 1053-1062), we have recovered cDNAs of spliced E2 mRNAs of human papillomavirus type 11 and additional mRNAs of type 11 and type 18 and determined the utilization of open reading frames (ORFs) in the DNA sequences. The recovery of cDNA copies of messages with splice sites identical to those previously described strongly suggests that the newly characterized splice donors and acceptors are also authentic. The HPV-11 E2 cDNA contains the intact E6 and E7 ORFs and the beginning of the El ORF in the first axon, which is then spliced from nt 847 to the second exon at nt 2622, 100 nucleotides upstream from the initiation codon for the E2 ORF. The initiation codon in the El ORF is followed by four additional in-frame AUG triplets and an in-frame termination codon positioned 30 nucleotides upstream from the initiation codon for the E2 protein. The authenticity of this putative E2 cDNA was shown by its ability to provide enhancer transactivating activity in chloramphenicol acetyltransferase (CAT) assays in several cell lines. A mutation in the genomic DNA at this splice acceptor site eliminates its activity, demonstrating that the splice is essential for the expression of the E2 protein. We conclude that the translation of the HPV-11 E2 protein requires internal initiation.

Original languageEnglish (US)
Pages (from-to)468-477
Number of pages10
JournalVirology
Volume172
Issue number2
DOIs
StatePublished - 1989

Fingerprint

Human papillomavirus 11
Retroviridae
Open Reading Frames
Complementary DNA
Initiator Codon
Messenger RNA
Genes
Nucleotides
Moloney murine leukemia virus
RNA Splice Sites
Chloramphenicol O-Acetyltransferase
Terminator Codon
Axons
Organism Cloning
Exons
Proteins
Epithelium
Cell Culture Techniques
Viruses
Cell Line

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Characterization of cDNAs of spliced HPV-11 E2 mRNA and other HPV mRNAs recovered via retrovirus-mediated gene transfer. / Rotenberg, Mitch 0.; Chiang, Cheng Ming; Ho, Maureen L.; Broker, Thomas R.; Chows, Louise T.

In: Virology, Vol. 172, No. 2, 1989, p. 468-477.

Research output: Contribution to journalArticle

Rotenberg, Mitch 0. ; Chiang, Cheng Ming ; Ho, Maureen L. ; Broker, Thomas R. ; Chows, Louise T. / Characterization of cDNAs of spliced HPV-11 E2 mRNA and other HPV mRNAs recovered via retrovirus-mediated gene transfer. In: Virology. 1989 ; Vol. 172, No. 2. pp. 468-477.
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