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Identification of the Human Papillomavirus Type 18 E6* and E6 Proteins in Nuclear Protein Fractions from Human Cervical Carcinoma Cells Grown in the Nude Mouse or in Vitro¹

Institut für Virusforschung, Deutsches Krebsforschungszentrum, D69 Heidelberg, [A. S. G., E. S.]; Universitáts-Frauenklinik, D6650 Homburg, [S. K., G. B.]; and European Molecular Biology Laboratory, D69 Heidelberg, [H. G., R. F.]; Federal Republic of Germany

We recently reported the transcription patterns of human papillomavirus (HPV) type 18 sequences in human cervical carcinoma cell lines. The open reading frames (ORFs) E6* and E6 represent the 5'-terminal cistrons in HPV18 mRNAs. ORF E6* was assumed to be specific for HPV types associated with genital carcinomas. To identify the predicted gene product, ORF E6* from a HeLa cDNA clone was expressed as an MS2 fusion protein in Escherichia coli. The C-terminal 23 amino acid residues were chemically synthesized. A panel of monoclonal antibodies was generated, recognizing E6* and E6* plus E6, respectively. In human cervical carcinoma cell lines grown in vitro these monoclonal antibodies specifically immunoprecipitate the putative M_r 17,000 and 18,000 HPV18 E6 proteins in nuclear protein fractions. In a HPV18 DNA containing human cervical carcinoma established in nude mice, these monoclonal antibodies specifically immunoprecipitate a polypeptide with a molecular weight of 6500 as predicted for the HPV18 ORF E6* gene product in a nuclear protein fraction.

¹ This work was supported by The Deutsche Forschungsgemeinschaft and by the Behringwerke AG, Marburg.

² Present address: Department of Biochemistry and Molecular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138. To whom requests for reprints should be addressed.

Received 12/ 4/87. Revised 3/ 1/88. Accepted 3/ 7/88.

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