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

在丝状真菌模式系统中性曲霉（Aspergillus nidulans）的营养生长中需要类化修饰，而COP9信号体（CSN）的去皮修饰仅在有性生命周期中必不可少。此外，CSN还需要光的控制和次级代谢的协调。CSN在酵母-2杂交系统中与DenA/Den1相互作用，DenA/Den1是真菌无性循环所需的另一种脱醛酶。我们将探讨CSN与DenA/1以及其他可能的相互作用伙伴（如cullins或两种真菌CandA/1蛋白）之间相互作用的功能。我们将特别强调DenA/1是否与整个CSN或仅与子复合物相互作用。标记的RubA/Nedd8在csnE/Csn5突变体中招募，但在cullin复合物和一些相关蛋白的野生型不同成分中不招募。我们将分析是否存在特定的CSN控制发育特异性真菌cullin复合物及其假定的底物。最后，我们将探讨在这种真菌性循环的果体形成过程中，是否存在Nedd8和SUMO偶联（或解偶联）途径之间的串音，因为SUMO基因的缺失会导致与无枝假单胞菌的去eddylase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.