Novel screens to identify genes regulating global chromatin structure during female meiotic prophase
Loh, Benjamin Jia Hui
During female meiotic prophase in many organisms, a specialized chromatin structure is formed in the oocyte nucleus. This structure is known as the karyosome, and has been proposed to be important for the formation of the female meiotic bipolar spindle. However, how the karyosome is formed and maintained is not very well understood. To identify proteins involved in the formation and maintenance of the karyosome, I carried out a cytological screen on a collection of 220 mutant fly lines for mutants that were defective in karyosome morphology. The screen identified 46 mutants on the X and 2nd chromosome with abnormal karyosomes. Genetic analysis of these 46 mutants, followed by molecular analysis of one mutant, identified SRPK (SR Protein Kinase) as a protein that is important for the proper formation of the karyosome. NHK-1 (Nucleosomal Histone Kinase 1) was previously identified as a protein that is essential for the formation of the karyosome via its phosphorylation of BAF (Barrier-to-Autointegration Factor). NHK-1 phosphorylation of BAF leads to the release of chromatin from the nuclear membrane, an essential step for the formation of the karyosome, however, the regulation of this process is unclear. In order to identify genes that interact with NHK-1, I carried out a genetic modifier screen using a semi-lethal allele of NHK-1, NHK-1trip. After screening a collection of 44 deficiencies located on the 2nd chromosome, I identified a genetic region (44B8-44D1) containing a gene that interacts with NHK-1 and, when gene dosage is halved, enhanced the semi-lethal phenotype of NHK-1trip.