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J Virol. 1994 November; 68(11): 7051-7059

Comparison of the properties of the E6 and E7 genes of low- and high-risk cutaneous papillomaviruses reveals strongly transforming and high Rb-binding activity for the E7 protein of the low-risk human papillomavirus type 1.

A Schmitt, J B Harry, B Rapp, F O Wettstein and T Iftner

Institut für Klinische und Molekulare Virologie, Erlangen, Germany.

ABSTRACT

A comparative analysis of different properties of the E6 and E7 proteins of high-risk and low-risk cutaneous papillomaviruses was performed. The corresponding genomic regions of human papillomavirus types 1 and 8 (HPV1 and HPV8) and of the cottontail rabbit papillomavirus (CRPV) were cloned into the eucaryotic expression vector pZipNeo-SV(X)-1 and into vectors for in vitro transcription and translation. With the help of these vectors, the individual proteins were investigated for their ability to transform C127 and NIH 3T3 rodent fibroblasts, bind the Rb protein in vitro, transactivate the adenovirus E2 promoter, and cooperate in the immortalization of primary human keratinocytes. Expression vectors for HPV16 E6 and E7 were used as a positive control. A highly transformed phenotype could be observed with rodent cell lines expressing HPV8 E6, HPV16 E6 and E7, and, surprisingly, HPV1 E7. In contrast, no transformation was detected with CRPV long E6 and HPV8 E7, whereas cells expressing HPV1 E6 and CRPV short E6 exhibited a weakly transformed phenotype. Although neither CRPV E6 nor CRPV E7 caused morphological transformation of C127 cells, CRPV E6 was able to induce anchorage-independent growth in both rodent cell lines, whereas CRPV E7 led to high cloning efficiencies only in NIH 3T3 cells. The in vitro Rb-binding affinities relative to that of HPV 16 E7 were 66% for HPV1 E7, 34% for HPV8 E7, and 11% for CRPV E7. In spite of its high Rb-binding affinity, HPV1 E7 did not trans activate the adenovirus E2 promoter, whereas HPV8 E7 and CRPV E7 showed low activities. Complementation studies in primary human keratinocytes revealed a weak immortalizing potential for HPV8 E7 and indicated a low degree of cooperativity between CRPV E7 and CRPV or HPV16 E6.

J Virol. 1994 November; 68(11): 7051-7059




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