Structural studies of the Golgi reassembly and stacking protein from S. cerevisiae and its golgin

Abstract

The current research project is an extension of the FAPESP PhD scholarship hold by Natália Fontana and is part of a FAPESP/BBSRC grant (Proc. FAPESP 2015/50366-7) aimed at supporting a formal collaboration between our group and Prof. Anthony Watts' group at the University of Oxford. The main part of the project is thus related to the student's ongoing PhD project in Brazil, which involves the Golgi Reassembly and Stacking Protein from Saccharomyces cereviseae (ScGRASP - Grh1). We previously showed that ScGRASP shares many of the uncommon features of the molten globule state (manuscript submitted to Scientific Reports). Furthermore, we proved that Grh1 forms amyloid-like fibrils, and we have enough evidence to show that it also happens in vivo, when the cell is under stress conditions. These results impact the way we think about GRASP functions in the cell and give new insights on the role of GRASP in the mechanisms of unconventional protein secretion. We intend to expand this scenario by getting higher-resolution data on ScGRASP fibril structure and also more information about its partner, the Golgin BUG1. The complex Grh1-BUG1 is fundamental for membrane tethering and secretion trough Compartments for nonconventional Secretion (CUPs) and little information is currently available about it. Our goal will be achieved by using Prof. Watts' expertise in both magnetic resonance and membrane studies, and of course all the infrastructure of the Department of Biochemistry in Oxford. The methods the student will be trained at Oxford will further improve our group's capacity to data analyses of non-conventional proteins, therefore positively affecting all the ongoing projects in our group in Brazil. (AU)