Sentrozom kümesi üzerine odakli Nek2a kinaz hedeflerinin incelenmesi

Cancer cells, unlike normal cells, usually have extra centrosomes, which form multipolar spindles (MPS) and cause cell death. Nonetheless, they divide successfully and avoid the lethal implications of uneven genetic material segregation by clustering their extra centrosomes into two poles. Nek2A is a mitotic kinase that regulates a variety of mitotic events. In this study, we demonstrate that while reduction of Nek2A activity via knock-out, silencing or using specific inhibitors favours centrosome clustering, its overexpression unclusters extra centrosomes making cancer cells vulnerable to cell death. Interestingly, none of the centrosomal targets of Nek2 (C-Nap1, Rootletin or Gas2L1) or its targets that can induce genomic instability (TRF1 or HEC1) appeared to responsible for its action on clustering, suggesting that other targets may be involved in this process. To investigate whether the effect of Nek2A on MPS is a novel pathway or acted in concert with other known unclustering factors, we tested HSET and NuMA. Our findings revealed that HSET's unclustering activity was complementary to NEK2A, implying an independent process, whereas the suppression of NuMA could reverse this effect. We also performed TurboID proximity labelling analysis, uncovering several potential Nek2A targets that were either situated at the centrosome or along microtubules, including NuMA. Although NuMA was in proximity, it did not co-IP with Nek2A. Intriguingly, we identified KIF2C as a new interaction partner of Nek2A and our subsequent analysis indicated that silencing it attenuated Nek2A activity on centrosome clustering. In conclusion, Nek2A's role in centrosome clustering and the identification of new interaction partners like KIF2C provides a deeper comprehension of cancer cell biology and may offer innovative avenues for targeted cancer therapy.