Insertional inactivation of the p53 gene during friend leukemia: a new strategy for identifying tumor suppressor genes

Abstract

Rearrangements within the p53 gene are observed in a high proportion of independent erythroleukemic cell lines derived from the spleens of mice infected with Friend leukemia virus. The majority of cells with at least one rearranged p53 allele do not express detectable levels of p53 protein. Here, we show that in 4 out of 19 newly established erythroleukemic cell lines induced by infecting DBA mice with either the anemia (FV-A) or polycythemia (FV-P)-inducing strains of Friend virus, the p53 gene is rearranged as a result of integration of spleen focus-forming virus (SFFV). Integration of SFFV within the p53 gene resulted in inactivation of gene expression as determined by Western blot analysis. The sites of SFFV integration in the p53 gene were found, by Southern blot analysis and the polymerase chain reaction, to be localized in a 1-kbp region between introns 7 and 9. In addition, loss of the normal p53 allele was observed in three of the erythroleukemic cell lines that carried a rearranged p53 gene. Insertion of SFFV in these cell lines resulted in either the appearance of aberrant p53 transcripts or the complete lack of p53 expression. The results presented in this paper demonstrate that retroviral insertions can not only contribute to neoplastic transformation by activating dominant oncogenes but also by inactivating genes that normally function in a negative way to control cell growth. Thus, it may be possible to identify additional genes of this class by characterizing chromosomal sequences that are adjacent to common sites of retroviral integration in tumors.