Presence of catenated human papillomavirus type 16 episomes in a cervical carcinoma cell line.
Human papillomavirus (HPV) is frequently associated with cervical carcinoma and derived cell lines. In primary tissues of the carcinoma, the viral genome may be present in episomal or integrated configuration. In cell lines, however, only integrated HPV sequences have been reported. In this article, we describe the presence of episomal type 16 HPV (HPV16), demonstrated by electron microscopy and two-dimensional agarose gel electrophoresis, in a cervical carcinoma cell line, CC7T/VGH, established in 1980 in Taiwan. In CC7T/VGH, the HPV16 sequences are transcriptionally active, and at least three major HPV16 RNA species were detected in Northern blots. Results from restriction enzyme and S1 nuclease analysis suggest a composition of oligomeric HPV16 molecules in dimeric repeats. In addition, the HPV16 oligomers exist as catenated molecules of interlocking rings instead of concatemers. A monomeric copy of the HPV16 episome was cloned from a Hirt supernatant of CC7T/VGH by using a plasmid vector. Mapping and partial sequencing studies revealed an internal deletion of 163 base pairs within the L1 open reading frame. However, insertion of an A.C nucleotide pair at the deletion junction restored the otherwise frame-shifted L1 open reading frame. Two base transitions were also found within the E7 and the E1 open reading frames. Our findings suggest the need for closer examination for HPV episomal catenation in other cervical carcinoma cell lines as well as in primary carcinoma tissues of the uterine cervix and the anogenital tract. With CC7T/VGH, a way is now available for studies of many important aspects of the biology of HPV such as replication and gene expression of the extrachromosomal viral genome.
Choo
K B
KB
Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, Republic of China.
Cheung
W F
WF
Liew
L N
LN
Lee
H H
HH
Han
S H
SH
D001483:Base Sequence; D002277:Carcinoma / Q000235:genetics / Q000382:microbiology*; D004273:DNA, Neoplasm / Q000032:analysis*; D004279:DNA, Viral / Q000302:isolation & purification* / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D027383:Papillomaviridae / Q000302:isolation & purification*; D010957:Plasmids; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured / Q000382:microbiology; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000382:microbiology*
D004273:DNA, Neoplasm; D004279:DNA, Viral