Human papillomaviruses associated with epidermodysplasia verruciformis. II. Molecular cloning and biochemical characterization of human papillomavirus 3a, 8, 10, and 12 genomes

Abstract

The DNA of 4 human papillomaviruses (HPV) that were found in the benign lesions of 3 patients suffering from epidermodysplasia verruciformis were characterized. The flat wart-like lesions and the macular lesions of patient 1 contained 2 viruses, HPV-3a and HPV-8, respectively, whose genomes had previously been only partially characterized. The flat wart-like lesions of patient 2 and the macular lesions of patient 3 each contained a virus previously considered as belonging to types 3 and 5, respectively. These viruses are different from all of the HPV types so far characterized; they have tentatively been named HPV-10 and HPV-12. The HPV-3a, HPV-8 and HPV-12 DNA and the 2 SalI fragments of HPV-10 DNA (94.1 and 5.9% of the genome length) were cloned in Escherichia coli after having been inserted in plasmid pBR322. The cloned HPV genomes have similar sizes (.apprx. 7700 base pairs), but their guanine-plus-cytosine contents differ from 41.8% for HPV-12 DNA to 45.5% for HPV-3a DNA. The study of the sensitivity of the 4 HPV DNA to 14 restriction endonucleases permitted the construction of cleavage maps. Evidence for conserved restriction sites was found only for the HPV-3a and HPV-10 genomes since 5 of the 21 restriction sites localized in the HPV-3a DNA seem to be present also in the HPV-10 DNA. Hybridization experiments, performed in liquid phase at saturation, showed a 35% sequence homology between HPV-3a and HPV-10 DNA, 17-29% sequence homology among HPV-5, HPV-8 and HPV-12 DNA, almost no sequence homology between the HPV-3a or HPV-10 DNA and the other HPV DNA, and a weak homology between HPV-9 DNA and HPV-8 or HPV-12 DNA. Blot hybridization experiments showed no sequence homology between the HPV-3a, HPV-8 and HPV-12 DNA and the DNA of the HPV associated with skin warts (HPV-1a, HPV-2, HPV-4 and HPV-7) or with mucocutaneous and mucous membrane lesions (HPV-6b and HPV-11a, respectively). One exception was a weak sequence homology between the HPV-2 prototype and HPV-3a or HPV-10 DNA. Evidently, at least the following 6 HPV types are associated with epidermodysplasia verruciformis: HPV-3a and HPV-10, which are associated with flat warts in the general population, and HPV-5, HPV-8, HPV-9 and HPV-12, which are associated specifically with epidermodysplasia verruciformis.