NEK2 phosphorylation antagonizes the microtubule stabilizing activity of centrobin.

Centrobin was initially identified as a centrosome protein for centriole duplication. Centrobin is also detected outside the centrosome and involved in other cellular functions, such as spindle assembly. We previously reported that centrobin is a substrate of both NEK2 and PLK1, but it is not clear what functional properties of centrobin are regulated by two kinases. Here, we report that centrobin is involved in cell spreading, migration and microtubule stabilization in interphase cells. The NEK2-depleted cells looked spread with well-developed microtubule networks and migrated faster than the control cells. The microtubule stability in NEK2-depleted cells was higher than the control cells. However, the opposite was the case in centrobin-depleted cells. The opposite outcomes in NEK2- and centrobin-depleted cells suggest that NEK2 antagonizes biological functions of centrobin. We identified NEK2 phosphorylation sites within centrobin, which is distinct from the PLK1 phosphorylation sites. In fact, the phospho-resistant mutant of centrobin against NEK2 stabilized microtubule networks in vivo. Based on the results, we propose that NEK2 phosphorylation antagonizes the microtubule stabilizing activity of centrobin. Centrobin is a novel example that NEK2 and PLK1 independently phosphorylate a substrate and result in opposite outcomes in substrate function.