Selective compartmentalization of different mdm2 proteins within the nucleus.

Overexpression of the mdm2 protooncogene protein, which can lead to the inactivation of normal p53, has been observed in some human cancers. The mdm2 gene is positively regulated by p53, providing for a feedback loop in the control of both p53 and mdm2 activity. The expression of the mdm2 and p53 proteins in different non-small cell lung carcinoma (NSCLC) cell types harboring wild-type or mutant p53, or lacking p53 altogether, were investigated to determine whether a correlation exists between the expression of these two proteins. The mdm2 protein was expressed at very low levels in all NSCLC lines examined, regardless of the p53 status. To determine whether mdm2 could be induced by p53 in NSCLC, NSCLC cells were transfected with a recombinant adenovirus expressing high levels of wild-type p53. The highest levels of exogenous wild-type p53 were observed in p53-null H358 and H1299 cells and in H226b cells expressing endogenous wild-type p53 were observed in p53-null H358 and H1299 cells and in H226b cells expressing endogenous wild-type p53. In these cells, wild-type p53 induced the expression of 90/92K M(r) mdm2 proteins, as well as several faster-migrating mdm2-related species exhibiting relative mobilities of 76/78K, 57/59K, 46K, 28K, and 12K. Northern analyses of H358 and H1299 cells transfected with wild-type p53 showed that these cells expressed three species of mdm2 mRNA of 5.5, 4.6-3.8, and 2.1 Kb in size. Subcellular fractionation revealed that the 90/92K M(r) mdm2 protein species was localized to both the crude plasma membrane/cytoplasmic and nuclear fractions, and that the smaller mdm2 proteins associated selectively with different nuclear substructures. The 76/78K, 57/59K, and 46K Mr(r) mdm2 proteins may be derived by differential splicing of the 5.5 Kb mRNA, and their differential compartmentalization within the nucleus suggests that each has a distinct function, potentially in the regulation of p53 and other gene products.