A viral-cellular junction fragment from a human papillomavirus type 16-positive tumor is competent in transformation of NIH 3T3 cells.
A 4.4-kilobase DNA fragment (T4.4) from a human tumor (comprising part of the human papillomavirus type 16 E6 promoter; the E6, E7, and part of the E1 open reading frames; and cellular sequences) was found to be competent to fully transform NIH 3T3 cells. This competency resides in the whole hybrid DNA fragment, since the separate viral or cellular DNA sequences were not active. Abundant E6-E7 transcripts were found in the transformed cells. When the cellular fragments were substituted with polyadenylation sequences from polyomavirus or simian virus 40 DNA, little or no restoration of transforming activity was observed. In experiments in which an exogenous reporting gene, that for chloramphenicol acetyltransferase, was used, the possibility was excluded that the cellular flanking sequences act as a traditional enhancer; yet, when the cellular sequences were placed downstream of a chloramphenicol acetyltransferase expression vector (pSV2 CAT), activity of the reference gene was clearly enhanced. These results indicate that DNA containing human papillomavirus type 16 open reading frames E6 and E7 isolated from the genome of a human tumor has transforming potential, that this potential is realized when the viral DNA is joined to cellular sequences, and that the cellular sequences function in a more complex way than by simply providing polyadenylation signals.
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J Y
JY
Department of Pathology, New York University Medical Center, New York 10016.
Defendi
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D000818:Animals; D015152:Blotting, Northern; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D008175:Lung Neoplasms / Q000382:microbiology; D051379:Mice; D008819:Mice, Nude; D027383:Papillomaviridae / Q000235:genetics* / Q000472:pathogenicity; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000502:physiology
D004279:DNA, Viral; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase