Dr. Sklar has identified two previously unknown genes, termed JAZF1 and 2, which are fused in low grade endometrial stromal sarcomas (ESSs) as a consequence of a recurrent (7;17)(p15;q21) chromosomal translocation associated with these tumors. Both genes encode proteins containing zinc finger domains, and in JAZF1, the sequences of codons for two such domains closely resemble the sequence for two zinc fingers found in the transcription factor Sfplp, which has recently been implicated in the mechanism governing the G2/M checkpoint in the cell cycle of S. cervesiae. He proposes to characterize the JAZF1 and 2 genes in tumors and normal tissues through a series of studies, beginning with the completion of nucleotide sequence analysis of the amino terminal ends of the normal coding sequences, which have not yet been located. A wide variety of uterine and non-uterine sarcomas will be screened for the fusion transcript and mutations of the JAZF1 gene to determine the range of neoplasms containing abnormalities in the two genes and to answer a number of outstanding questions regarding the classification and histogenesis of uterine stromal tumors. Additionally, the ability of the JAZF1/JAZF2 fusion to induce malignant transformation will be investigated using both cultured cells and transgenic mice. Gene expression altered by the presence of the JAZF1/JAZF2 fusion will be analyzed by examining transcription profiles in cultured cells carry cDNA for the two genes transfected into bacteria. Interactions between the protein products of the two genes and other proteins will be explored using the yeast two-hybrid system. Phenotypic effects of the murine homologs for the JAZF1 and 2 will be assessed in knockout animals separately defective in both alleles. Finally, DNA binding sites for the two proteins will be sought and the sequences of these sites will be used in turn to identify genes transcriptionally regulated by JAZF1 and 2 in normal cells.
