Abstract 5029: A computational analysis of NEK10 and its novel protein-protein interaction with HspB1

Abstract The NEK kinase family of proteins consists of 11 serine/threonine kinases that participate in the disjunction of the centrosome, mitotic spindle assembly, and primary cilium formation. NEK10 is the most divergent member of the NEK family. It is unique in having a catalytic domain that is centrally positioned and flanked by two coiled-coil domains, while all the other NEKs have their catalytic domain near the N-terminus. Instead, NEK10 has four armadillo repeats of unexplored function in its N-terminus. NEK10 seems to play a key role in carcinogenesis and has been linked in melanoma, breast cancer, and a variety of ciliopathies. As part of our long-term goal to build interactomes of all the NEK members, we have previously reported data for known and predicted NEK10 interacting proteins, including novel protein-protein interactions such as HspB1 and MAP3K1, which have not been previously reported in the literature. In this study, we focused on understanding the molecular mechanism underlying NEK10’s interaction with HspB1 and its functional consequences. HspB1 is involved in various cellular functions such as maintaining cytoskeletal integrity and cell death. It is also a well-studied cancer protein and contains a highly conserved α-crystallin domain flanked by disordered N- and C-terminal domains. Many proteins are targeted by HspB1 to promote resistance to cell death, and malignant phenotypes; high levels of HspB1 have been identified in many cancer stem cells, such as those from lung and breast cancers. We used state-of-the-art computational approaches to model and characterize the full-length NEK10 as well as HspB1 protein. Our results offer robust full-length three-dimensional models of NEK10 and HspB1 and their biophysical characterization. In addition, we delineate the role of the armadillo repeats that are unique to NEK10 in the NEK family, in its interaction with HspB1. Our docking analysis shows that H301 of NEK10 located within its Armadillo Repeats forms a salt bridge with I179 of HspB1. Our results, therefore, suggest a scenario of Nek10 interaction with HspB1 using its ARM motif, which frees the catalytic domain for auto-phosphorylation and/or phosphorylation of other targets. Since HspB1 regulates the architecture of the cellular microtubular network (MT) by facilitating the formation of non-centrosomal MT, we speculate that the interaction of NEK10 with HspB1 plays a role in centrosome dynamics. Overall, this study shows novel and intriguing information about NEK10’s structure-function relationships, especially in the context of its interaction with HspB1. Furthermore, it establishes the framework for elucidating the detailed molecular mechanisms of NEK10 interactions with other proteins to further explore its potential as a therapeutic target. Citation Format: Andriele Silva, Shaneen Singh. A computational analysis of NEK10 and its novel protein-protein interaction with HspB1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5029.