Ayscough, Kathryn Rachel; (1993) Morphological analysis of the Golgi apparatus in Schizosaccharomyces pombe. Doctoral thesis (Ph.D.), University College London (United Kingdom).

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Abstract

The most striking, morphological aspect of the Golgi apparatus is its stack of cisternae. How this distinct structure relates to its known function in the processing of secretory proteins, is however, not known. During the course of my work I have made an intensive investigation of the morphology of the Golgi apparatus in Schizosaccharomyces pombe. This yeast contains stacks of membranes resembling the Golgi stacks of mammalian cells suggesting that certain fundamental processes underlying formation of the stack may be conserved. I have demonstrated that the Golgi cisternae can be reversibly unstacked in vivo and that stacking required intact cytoplasmic microtubules. This was revealed by use of microtubule disrupting drugs and tubulin mutants. The functional consequences of this morphological change were addressed. Inhibition of protein synthesis and a mutation in a gene product required early in the secretory pathway resulted in breakdown of the Golgi apparatus. The action of Brefeldin A was investigated and resembled its effect in mammalian cells with Golgi disruption and redistribution to the ER. Finally, specific cross reactivity with an antibody to mammalian Golgi structures showed that some epitopes have been conserved over the evolutionary period of divergence from S. pombe to mammalian cells. This study has highlighted many aspects of the Golgi structure and further research at a genetic level would reveal molecules involved in the maintenance of the stack. Such work would in turn increase our understanding of the role of cisternal stacking.

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