Specific Cooperation Between Imp-α2 and Imp-β/Ketel in Spindle Assembly DuringDrosophilaEarly Nuclear Divisions

Abstract

AbstractThe multifunctional factors Imp-α and Imp-β are involved in nuclear protein import, mitotic spindle dynamics, and nuclear membrane formation. Furthermore, each of the three members of the Imp-α family exerts distinct tasks during development. In Drosophila melanogaster, the imp-α2 gene is critical during oogenesis for ring canal assembly; specific mutations, which allow oogenesis to proceed normally, were found to block early embryonic mitosis. Here, we show that imp-α2 and imp-β genetically interact during early embryonic development, and we characterize the pattern of defects affecting mitosis in embryos laid by heterozygous imp-α2D14 and imp-βKetRE34 females. Embryonic development is arrested in these embryos but is unaffected in combinations between imp-βKetRE34 and null mutations in imp-α1 or imp-α3. Furthermore, the imp-α2D14/imp-βKetRE34 interaction could only be rescued by an imp-α2 transgene, albeit not imp-α1 or imp-α3, showing the exclusive imp-α2 function with imp-β. Use of transgenes carrying modifications in the major Imp-α2 domains showed the critical requirement of the nuclear localization signal binding (NLSB) site in this process. In the mutant embryos, we found metaphase-arrested mitoses made of enlarged spindles, suggesting an unrestrained activity of factors promoting spindle assembly. In accordance with this, we found that Imp-βKetRE34 and Imp-βKetD bind a high level of RanGTP/GDP, and a deletion decreasing RanGTP level suppresses the imp-βKetRE34 phenotype. These data suggest that a fine balance among Imp-α2, Imp-β, RanGTP, and the NLS cargos is critical for mitotic progression during early embryonic development.