Molecular cloning and characterization of endogenous SV40 dna from human HBL-100 cells

Claude Saint-Ruf 1 P. Nardeux 1 J. Cebrian 2 M. Lacasa 2 C. Lavialle 1 Roland Cassingena 1
1 UPR6 CNRS - Laboratoire d'Immunologie [Villejuif]
2 UPR2 CNRS - Laboratoire d'Immunologie [Villejuif]
Abstract : The human HBL-100 cell line harbours SV40 DNA integrated in tandem at a unique site. The SV40 T-antigen expressed in these cells is defective in a function essential to the replication of the viral genome. The integrated SV40 sequences were molecularly cloned in a bacteriophage, and a subclone (plasmid pSVHBI) containing a complete SV40 DNA was isolated. As compared to SV40 wild-type strain 776, sequence analysis of pSVHBI early region revealed the presence of several DNA alterations. Among these, a point mutation at position 3199, predicting a change at amino-acid 540 of arginine to isoleucine, was shown by marker rescue to be responsible for the deficiency of T-antigen. This novel mutation further delimits one of the T-antigen domains involved in SV40 DNA replication. Transfection experiments demonstrated that the transforming activity of the SV40 genome from HBL-100 cells is still preserved. Moreover, several transformed human cell clones thus obtained could be permanently established in culture.
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Claude Saint-Ruf, P. Nardeux, J. Cebrian, M. Lacasa, C. Lavialle, et al.. Molecular cloning and characterization of endogenous SV40 dna from human HBL-100 cells. International Journal of Cancer, Wiley, 1989, 44 (2), pp.367-372. ⟨10.1002/ijc.2910440230⟩. ⟨inserm-02179369⟩

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