Identification and characterization of NEDD8-modified proteins and deneddylases of the filamentous fungs Aspergillus nidulans

丝状真菌模型系统Nidulans的营养生长需要去核化，而COP9信号体(CSN)去核化是有性生命周期所必需的。此外，CSN对于光控制和次生代谢的协调也是必需的。CSN在酵母-2-杂交系统中与DenA/Den1相互作用，DenA/Den1是真菌无性循环所必需的另一种脱氢酶。我们将探索CSN和Dena/1之间的相互作用以及其他可能的相互作用伙伴，如Cullins或两个真菌Canda/1蛋白。我们将特别强调Dena/1是与整个CSN相互作用，还是仅与亚复合体相互作用。标记的Ruba/Nedd8在csnE/csn5突变体中招募，但不在野生型cullin复合体的不同成分和几个相关蛋白中招募。我们将分析是否存在发育特异性真菌cullin复合体及其假定底物的特异性CSN控制。最后，我们将探索在这种真菌有性周期的子实体形成过程中，Nedd8和相扑接合(或去共轭)途径之间是否存在串扰，因为相扑基因的缺失导致了与脱氧核酸酶缺陷突变体相似的发育表型。

Neddylation is required for vegetative growth of the filamentous fungal model system Aspergillus nidulans, whereas deneddylation by the COP9 signalsome (CSN) is only essential for the sexual life cycle. In addition, CSN is required for light control and coordination of secondary metabolism. CSN interacts in a yeast-2-hybrid system to DenA/Den1, another deneddylase which is required for the asexual cycle of the fungus. We will explore the function of the interaction between CSN and DenA/1 and other putative interaction partners as cullins or the two fungal CandA/1 proteins. We will especiallyemphasize whether DenA/1 interacts to the entire CSN or only to subcomplexes. A tagged RubA/Nedd8 recruits in csnE/Csn5 mutants but not in wildtype distinct components of cullin complexes and several associated proteins. We will analyse whether there is a specific CSN control of developmental-specific fungal cullin complexes and their putative substrates. Finally we will explore whether there is a crosstalk between the Nedd8 and the SUMO conjugation (or deconjugation) pathway during fruit body formation of the sexual cycle of this fungus, because a deletion of the gene for SUMO results in a similar developmental phenotype as a deneddylase deficient mutant of A. nidulans.