Close cooperation between Semi1 and Semi2 proteins is essential for pronuclear positioning in Tetrahymena thermophila.

During sexual reproduction in the ciliate Tetrahymena thermophila, meiosis occurs in the germline micronucleus, resulting in the formation of four haploid micronuclei. Of these, only one is selected to evade autophagy, and subsequently migrates to the membrane junction with the partner cell for reciprocal pronuclear exchange. We previously demonstrated that the transmembrane protein Semi1 is essential for this nuclear migration. Semi1 is specifically expressed in mating cells and localizes to the periphery of the selected nucleus. Loss of Semi1 disrupts nuclear attachment to the junction, leading to infertility. However, the mechanism by which Semi1 positions the nucleus at the junction remains unclear. Here, we report that the Semi1-interacting protein, Semi2, is also necessary for proper nuclear positioning. Deletion of Semi2 results in the same nuclear mislocalization phenotype and infertility observed in Semi1 mutant cells. Semi2 colocalizes with Semi1, but in the absence of Semi1, Semi2 fails to exhibit perinuclear localization. The selected nucleus anchors to microtubules prior to migration, a process dependent on both Semi1 and Semi2. We propose a model in which Semi1 recruits Semi2 to the selected nucleus, facilitating the interaction between the nucleus and microtubules required for proper nuclear positioning at the membrane junction.