Although the gene responsible for multiple endocrine neoplasia, type 1 (MEN1) has been identified recently, the function of its gene product, menin, is not known. To examine menin's biological role, we created an N-terminal tagged fusion protein to follow the distribution of menin in the cell. In all cell lines tested, menin was found both in the nucleus and the cytoplasm, but its localization was dependent on the phase of the cell cycle; during a nondividing phase, menin was found in the nucleus; during and immediately after cell division, it was found in the cytoplasm. To confirm the cellular localization seen with the N-terminal tagged protein, we developed and purified peptide-specific antibodies. One of these antibodies (NCI 624), which recognizes a domain (aa 383-395) of menin, was used in immunofluorescence studies to corroborate the N-terminal tagging results. Further confirmation of menin localization was obtained in a pituitary tumor cell line derived from a familial MEN1 patient, which contained a mixed cell population with either none, or one functional copy of the MEN1 gene. Our results indicate that menin functions principally as a nuclear protein but may be found in the cytoplasm during cell division. (Lab Invest 1999, 79:301-310).