Accumulation of RNA homologous to human papillomavirus type 16 open reading frames in genital precancers

Crum, Christopher P.; Gj, Nuovo; Friedman, Daniel J.; Silverstein, Saul

doi:10.1128/jvi.62.1.84-90.1988

Cited by 88 publications

(42 citation statements)

References 36 publications

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“…High-risk HPVs are etiologic agents of anogenital tumors, including cervical cancers (51). The late viral functions of vegetative viral genome amplification, late gene expression, and virion morphogenesis are restricted to the upper, differentiated keratinocytes of the epithelium (4,8,16,17,30,42). The organotypic or raft culture system has permitted the study of complete vegetative life cycles of HPVs in vitro (24,30,31,35).…”

Section: Discussionmentioning

confidence: 99%

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Temporal Usage of Multiple Promoters during the Life Cycle of Human Papillomavirus Type 31b

Ozbun

Meyers

1998

J Virol

103

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The life cycles of human papillomaviruses (HPVs) are dependent upon the differentiation of the epithelial cells they infect. HPV type 31b (HPV31b) virions can be purified following the growth of a latently HPV-infected cell line (CIN-612 9E) in the organotypic or raft system. Treatment of the CIN-612 9E raft tissues with protein kinase C (PKC) activators is required for upregulation of late gene expression and efficient production of virions. We employed the raft culture system to study the temporal usage of HPV31b promoters during the viral life cycle. We compared monolayer cultures of CIN-612 9E cells, untreated CIN-612 9E raft tissues, and PKC-induced CIN-612 9E raft tissues harvested at various time points during epithelial differentiation. We found that the HPV31b major early promoter precisely maps to nucleotide (nt) 99 (P99). A transcriptional start site for both early and late gene transcripts mapped upstream of P99 at nt 77 (P77). The P77 and P99 promoters were used constitutively throughout the HPV31b life cycle; however, initiation from P99 was much stronger than from P77. Mapping of the differentiation-induced P742 promoter revealed multiple start sites. These start sites were difficult to detect in monolayer cultures, were induced in untreated rafts, and were greatest in PKC-induced raft tissues at 8 to 12 days. A constitutively active promoter, P3320, was also defined and is responsible for the transcription of unspliced and spliced RNAs containing E5a, E5b, L2, and L1 open reading frames.

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Section: Discussionmentioning

confidence: 99%

“…The genomic organization is highly conserved among HPVs, and the life cycles of the viruses are tightly linked to the differentiation state of the infected cells (8,30,42,45). The virions encapsidate a circular DNA molecule containing six to eight early open reading frames (ORFs) and two late ORFs.…”

mentioning

confidence: 99%

Temporal Usage of Multiple Promoters during the Life Cycle of Human Papillomavirus Type 31b

Ozbun

Meyers

1998

J Virol

103

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“…The detection of late transcripts in HPKII cells and invasive carcinomas containing integrated genomes of HPV 16 implies that HPV 16 late gene transcription is not completely limited to terminally differentiated cells as previously suggested (Crum et al, 1988), and could occur from integrated genomes provided the respective region was preserved.…”

Section: Rna Pcr Analysis Of Hpv 16 Transcripts In Biopv Specimens Frmentioning

confidence: 99%

“…In situ hybridization studies in CIN biopsies demonstrated that the most abundant hybridization signals occurred from the E4-ES orfs, with weaker signals from the E6-E7 orfs and the 5' portion of the E2 orf (Crum et al, 1988). Northern blot analyses with subgenomic DNA probes revealed 4 types of mRNA which represented all the orfs of the early region (Shirasawa et al, 1988).…”

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confidence: 99%

Expression and splicing patterns of human papillomavirus type‐16 mrnas in pre‐cancerous lesions and carcinomas of the cervix, in human keratinocytes immortalized by HPV 16, and in cell lines established from cervical cancers

Sherman

Alloul

Golan

et al. 1992

Intl Journal of Cancer

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We have analysed the splicing patterns of human papillomavirus (HPV) type-16 mRNAs in a human epithelial cell line immortalized by HPV 16 (HPKII), in cell lines established from cervical carcinomas (SiHa and CaSki) and in pre-invasive and invasive carcinomas of the cervix. The presence of mRNA species previously described, which could encode the E6, E6I, E6II, E6III, E7, E2, E2C, E4, E5 and L1 proteins, was determined, using the RNA polymerase chain reaction (PCR) technique with primers that flank unique splice sites. The state of the viral DNA in the tumor biopsies was established by Southern blot analysis. The various HPV 16 transcripts could be detected in cell lines and in tumor biopsies. The size of the RNA PCR products were in agreement with the previously mapped splice sites. The full range of transcripts was revealed in the HPKII cell line and in a number of pre-invasive carcinomas. Messenger RNAs which could encode the E6III, E4 and E5 proteins were most prevalent in all types of tumor. The overall results of DNA and RNA analyses in cell lines and tumor specimens indicate that (1) expression of either of the early or late transcripts studied is not specifically related to (a) tumor stage or (b) the physical state of the viral genome; and (2) alterations in the splicing patterns of HPV 16 transcripts may not be involved in tumor progression.

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“…Therefore, the study of tissue structure of HPVs without DNA extraction is important. Immunohistochemistry and in situ hybridization are therefore used [Ferenczy et al, 1981;Jenkins et al, 1987;Crum et al, 1988;Syrjanen and Syrjanen, 1986;Stoler and Broker, 19861. In this report, the presence of HPV and the genome of HPV is described systematically in benign neoplasia, condyloma acuminatum, and dysplasia and in malignant neoplasia, intraepithelial carcinoma, and invasive carcinoma.…”

Section: Introductionmentioning

confidence: 99%

Human papillomavirus infection of the uterine cervix analyzed by nonisotopic in situ hybridization

Hara

Tuchida

Nakamura

et al. 1990

Journal of Medical Virology

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Some types of human papillomaviruses (HPVs) have been suggested to be strongly related to uterine cervical carcinoma. An attempt to detect these in formalin-fixed, paraffin-embedded sections was made by either immunohistochemical or by in situ hybridization. Anticapsid protein of bovine papillomavirus antibody labeled with peroxidase was used for immunohistochemistry, and biotin was used instead of radioisotopes to label probes for in situ hybridization, which resulted in low background and a rapid procedure. Condylomatous changes were stained immunochemically with this antibody even in invasive carcinoma, whereas the carcinoma itself was not stained. Direct correlation was demonstrated by in situ hybridization between the HPV genome and histopathological structure, which was impossible by Southern or dot hybridization. HPV DNAs were detected in the nuclei of koilocytes and dyskeratinocytes of condylomata and dysplasias. Furthermore, hybridization signals of HPV DNAs in basal and parabasal cells suggested that HPV infection had already begun in the basal cells. In the case of malignant neoplasia accompanied by dysplasia, the same type of HPV was detected both in the malignant neoplasia and accompanying dysplasia. In one case of intraepithelial carcinoma, the very small focus of carcinoma just arisen in the cells of dysplasia was identified, and both were positive for HPV 18. This fact supports the suggestion that the carcinoma arises in dysplasia. Invasive carcinomas were classified further into keratinized, large-cell nonkeratinized, and small-cell nonkeratinized types, and the positive frequency for HPV 16 decreased as the differentiation of the carcinoma decreased. In the case of keratinized type of invasive carcinoma, strong hybridization signals were prominent around the malignant pearl formation.

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Accumulation of RNA homologous to human papillomavirus type 16 open reading frames in genital precancers

Cited by 88 publications

References 36 publications

Temporal Usage of Multiple Promoters during the Life Cycle of Human Papillomavirus Type 31b

Temporal Usage of Multiple Promoters during the Life Cycle of Human Papillomavirus Type 31b

Expression and splicing patterns of human papillomavirus type‐16 mrnas in pre‐cancerous lesions and carcinomas of the cervix, in human keratinocytes immortalized by HPV 16, and in cell lines established from cervical cancers

Human papillomavirus infection of the uterine cervix analyzed by nonisotopic in situ hybridization

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