The role of Arf1 in maintaining Golgi structure and function

Daniele, Tiziana (2004). The role of Arf1 in maintaining Golgi structure and function. PhD thesis The Open University.



Despite the early recognition of the importance of Arf1 and its multiple effectors in the regulation of both the Golgi complex ultrastructure and function, the mechanisms by which Arf1 exerts its effects on the Golgi apparatus architecture and on intracellular transport are still poorly understood. Indeed, there is a gap between what is known on the interaction between Arfl1 and at least some of its effectors at the sub-molecular level, and the understanding of the relative contribution of each of the effectors to the final effect of Arf1 on the Golgi complex.

The aims of the present study were to determine the fine distribution of Arf1 in the Golgi apparatus, to assess which processes depend on Arf1 activity and/or GTP hydrolysis on Arf1 and to clarify the relative roles of at least two Arf1 effectors (PLD1 and coatomer) in the maintenance of the structure of the Golgi complex and membrane trafficking.

To this purpose, multiple approaches have been used targeting either Arf1 itself (dominant positive and dominant negative mutants) or directly Arf1 effectors (such as coatomer and PLD1). In all cases, a detailed analysis of different steps of transport and of the ultrastructure of the Golgi apparatus has been performed.

The main findings of this work include the definition of Arf1 distribution in the Golgi complex sub-compartments, the demonstration that the roles of coatomer in the regulation of ER-to-Golgi transport and of the architecture of the Golgi apparatus can be dissociated both functionally and kinetically, the assessment of the lack of any role of PLD in the modulation of both the structure and the function of the Golgi complex and, finally, the uncovering of an unanticipated role of Arf1 in TGN-to-PM traffic and the identification of two novel effectors of Arf1, acting in the generation of constitutive post-Golgi carriers.

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