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Proteolytic cleavage of the Three rows protein (THR) subunit during anaphase has been found to limit Drosophila separase function. THR cleavage is especially important during the process of cellularization, which is after completion of the last syncytial mitosis of early embryogenesis. This suggests that Drosophila separase has targets in addition to cohesin subunits.

Drosophila Separase has been found to be necessary for sister chromatid separation and it binds to THR and PIM, which associate in vivo. The Drosophila SSE sequence is very diverged. Some features conserved in homologs from trypanosomatids to vertebrates are not seen in it.

Degradation of Drosophila Pimples (PIM) protein has been found to regulate sister chromatid separation in mitosis. The nonconserved Drosophila genes, pimples and three rows, both necessary specifically for sister chromatid separation in mitosis, have been identified. PIM shares extensive functional similarity with securin proteins and with Cut2 from fission yeast an is found in a comlex with Three rows protein.

Cyclin B3 and Cyclin B are necessary for female fertility but neither is needed for mitosis in Drosophila. Cyclin B3 is found in proliferating cells during mitosis but cannot be detected in Drosophila embryos' tissues during endoreduplication. Cyclin B3 becomes degraded quickly during mitosis. Cyclin B3 is a nuclear protein, unlike Cyclins A and B, with which it cooperates in regulating mitosis.

A study was conducted to understand the Geminin protein oscillation in endoreplicating salivary glands of Drosophila. The results showed that APC/C activity is required in endoreplicating cells to mediate Geminin oscillation and stabilize the Geminin protein and block endocycle progression.

Study is conducted in which a genetic loci is screened that modify the aberrant phenotypes caused by over expression of regulatory separase complex subunits Pimples/securin and Three rows in Drosophila. An interacting gene is found to encode a constitutive centromere protein.