To unravel the interaction between Nup155 and Nup93 via structural studies

Abstract

The nuclear pore complex is a necessary protein assembly in the cell that regulates the trafficking between the nucleus and cytoplasm. This megadalton assembly consists of multiple subunits known as nucleoporins, which assemble themselves to form submodules. Owing to the huge size and technical limitations, the basic structural architecture and functional understanding of this complex is limited. The other way to comprehend the function of the nuclear pore complex is to delineate the interactions among individual proteins and their subcomplexes and hence, study their structure. In this study we investigated the interaction between the two critical proteins of the inner ring - Nup93 and Nup155. Various domains of these two proteins were reconstituted in-vitro using bacterial expression system. Purification optimisation was performed to further stabilise these interactions and obtain high yield. From this study, we were able to reconstitute the domains of Nup155 with its interacting partner Nup93 and also collected cryo-EM data for the complex. We performed some initial data processing, from which we have concluded that the quality of the data is good enough, and can be used for further processing. In future, we will be aiming for a high resolution structure of the Nup155.Nup93 complex by processing the data we have collected. The high-resolution structure of the complex can be used as a small piece in the big LEGO puzzle, which will ultimately contribute to better understanding of the NPC.