An investigation of the functional role of GRAB in the context of Rab3, Rab8 and Rab11

Abstract

Rab GTPases are the largest family of the Ras superfamily and are key regulators of membrane trafficking within the cell. There are over 60 members of the Rab family which localise to specific membrane compartments and interact with effector proteins to regulate membrane trafficking processes, such as vesicle formation, vesicle trafficking within the cell and fusion with an acceptor compartment. Multiple effector proteins have been identified for many Rabs, some of which can interact with more than one Rab to link their function at a specific membrane location or to link them together in a Rab activation cascade. Rabin8 is one such protein which is an effector for Rab11a and a Guanine nucleotide Exchange Factor (GEF) for Rab8a. Rabin8 participates in a conserved Rab activation cascade which is critical in the formation of primary cilia. Data presented in this thesis has shown that GRAB interacts with Rab3a, Rab8a, Rab11a and Rab11b in a nucleotide dependent manner. Furthermore, the minimal interacting regionbetween these proteins has been investigated. The functional outcome of GRAB knockdown has also been examined and data in this thesis highlights the phenotypic outcome.