Structure and function of SV 40 large T antigen: communication between functional domains.
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The mutagenic effect of papovavirus SV40 on human cells could be demonstrated by reversion of HPRT deficiency in Lesch-Nyhan fibroblasts transformed by the virus. SV40 seems to induce different gene mutations in individually selected cell clones, as was clearly shown by the respective HPRT enzyme properties. The consequences of our results for use of SV40-derived vectors in gene substitution experiments are discussed.
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Co-culturing with origin-defective SV40 DNA-injected transformants, we succeeded in activating the limited growth potential of late-phase-II fibroblasts from patients with Werner's syndrome (WS cells). Residual life spans of three WS cell lines increased 2.3-5.6 fold. Co-culture with normal IMR-90 cells showed variable effects on different WS cell lines.
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A human fibroblastic cell line transformed by the SV40-T antigen sequence and continuously cultured for 7 months displayed large periodic variations in cell proliferation. This contrasted with other characteristics of this cell line that remained constant: mosaic cell shape, absence of cell contact inhibition, and predominance of a hypodiploid population. Similar fluctuations in proliferative capacity were also found during the long-term growth of a transformed but nonimmortalized human fibroblastic line prior to senescence, and in the established hamster fibroblastic Nil cell line. This growth pattern suggests a recurrent stimulation of growth in these three transformed cell lines. The proliferation pattern from cultured transformed cells may thus be complex and requires further investigation. These variations presumably influence major cell functions. This observation has important implications for the analysis of data from such cell lines.
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