DNA sequences derived from the genome of the papillomavirus HPV3

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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536 2372, 536 2432, 935 77, 935 78, C07H 2102, C07H 2104

Patent

active

056564236

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The invention relates to specific DNA sequences derived from the genome of the papillomavirus HPV39, including the sequence corresponding to its entire genome as well as recombinant DNAs, in particular vectors containing these DNA sequences. The invention also relates to the cell cultures transformed with the said recombinant DNAs under conditions making it possible for them to express the corresponding sequences derived from the HPV39 genome in the form of the corresponding proteins. Finally, the invention relates to the purified proteins obtained from these cell cultures. Finally, the invention relates to the production of the immunogenic compositions containing such proteins or protein fragments.


BACKGROUND OF THE INVENTION

The invention is based on the discovery of specific sequences present in HPV39, sequences which constitute its originality and which make possible particularly discriminating detection of papillomaviruses of the HPV39 type. These sequences or fragments of these sequences can be used for constituting particularly sensitive hybridization probes, in particular, primers which make possible analyses by the so-called PCR method. Before proceeding further with the description of these sequences or sequence fragments, it is proposed to make a brief review of the state of the art and, then, to provide a detailed description of the genome of HPV39.
Among the 60 if not more different types of human papillomaviruses, some are associated with neoplasias or with carcinomas of the genital apparatus (1). The DNA of HPV16 and HPV18 were detected in 50% and 20% of the biopsies of cervical, vulvar or penile cancer (2,3). The HPV 31, 33, 35, 39 and 45 were encountered less frequently in such lesions (1,4). By using the DNA of HPV6 as a hybridization probe under conditions of low stringency, HPV39 was first cloned from biopsy samples of Bowenoid penile papules, which contain the viral DNA in its episomal form. On the other hand, the viral DNA was found to be integrated into the cellular genome of invasive carcinomas (5). In a recent study performed on 365 patients infected with HPV, the DNA of HPV39 was detected in 3.9% of the tissue samples.
The biological study of HPV has been hindered by the absence of a tissue culture system for in vitro viral propagation. The analysis of the sequence of a certain number of papillomavirus genomes (2, 3, 6-14) has provided the basis for the understanding of their genetic organization and their regulation, for the expression of individual genes and the generation of antisera, and for the evaluation of the phylogenetic relationships among the very many types of HPV.


BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Nucleotide sequence of the DNA strand of HPV39 analogous to the mRNA. Position 1 on the circular genome was determined by alignment with HPV16 and 18.
FIG. 2(a) and 2(b): Distribution of the start codons (bars above the line) and stop codons (bars below the line) in the three reading frames of the two strands of HPV39 DNA. FIG. 2(a) strand analogous to the mRNA, FIG. 2(b) complementary strand. The ORFs in (a) were identified by comparison with other types of HPV. The numbering is in agreement with that shown in FIG. 1.
FIG. 3: Principal characteristics of the non-coding region. The following sequence motifs of the NCR are shown from the nucleotide 7158 to the nucleotide 106: represents the palindrome of 12 bp specific for the papillomavirus (the second and third motifs are degenerate), represents the polyadenylation site, represents the TATA box, .rect-hollow. represents the presumed promoter element, represents the element of glucocorticoid response, represents the core enhancer sequence, represents the probable binding site of the nuclear factor I, represents the presumed binding site of the activating protein 1, represents the binding site for the factor associated with the papillomavirus enhancer.
FIG. 4: Comparison of the probable elements of glucocorticoid response (GREs) of HPV39 compared with the GREs of HPV16 and 18 and with a G

REFERENCES:
patent: 4983728 (1991-01-01), Herzog et al.
patent: 5182377 (1993-01-01), Manos et al.
patent: 5447839 (1995-09-01), Manos et al.
Beaudenon et al., Virology 161:374-384 (1987).
Reuter et al., J. Virol. 65(10):5564-5568 (Oct. 1991).
Volpers et al., Virology 181:419-423 (Mar. 1991).

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