Spot-1, a novel NLS-binding protein that interacts with p53 through a domain encoded by p(CA)n repeats

Nuclear Localization Signals (NLS) have been found to mediate the import of proteins into the nucleus. Proteins interacting directly with NLS control the subcellular localization of nucleophilic proteins. The p53 protein is spatially regulated throughout the cell cycle and this regulation has been shown to be dependent on the presence of its NLS sequences. We identified three novel cDNA clones that were isolated from an expression library because they encode polypeptides that bind a synthetic peptide containing the major NLS of p53 (NLS I). These clones were found to share a common domain encoded by p(CA)n repeats; a simple sequence length polymorphism (SSLP). THis is the first report where p(CA)n repeats were found to encode protein. One cDNA clone encodes a full length, 16 kDa protein, designated spot-1, that is represented in cells predominantly as oligomers. spot-1 interacts with the NLS I of p53 through its p(CA)n repeat. Cell fractionation and immunofluorescence analysis demonstrated that spot-1 is a nuclear protein which, in fibroblasts, co-localizes with p53.