Abstract
In Aspergillus nidulans, carbon catabolite repression (CCR) is mediated by the global repressor protein CreA. The deubiquitinating enzyme CreB is a component of the CCR network. Genetic interaction was confirmed using a strain containing complete loss-of-function alleles of both creA and creB. No direct physical interaction was identified between tagged versions of CreA and CreB. To identify any possible protein(s) that may form a bridge between CreA and CreB, we purified both proteins from mycelia grown in media that result in repression or derepression. The purified proteins were analysed by LC/MS and identified using MaxQuant and Mascot databases. For both CreA and CreB, 47 proteins were identified in repressing and derepressing conditions. Orthologues of the co-purified proteins were identified in S. cerevisiae and humans. Gene ontology analyses of A. nidulans proteins and yeast and human orthologues were performed. Functional annotation analysis revealed that proteins that preferentially interact with CreA in repressing conditions include histones and histone transcription regulator 3 (Hir3). Proteins interacting with CreB tend to be involved in cellular transportation and organization. Similar findings were obtained using yeast and human orthologues, although the yeast background generated a number of other biological processes involving Mig1p which were not present in the A. nidulans or human background analyses. Hir3 was present in repressing conditions for CreA and in both growth conditions for CreB, suggesting that Hir3, or proteins interacting with Hir3, could be a possible target of CreB.
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Acknowledgements
We thank Robin Lockington, Jai Denton, Adrian Hunter and Zhipeng Qu for helpful insights into the research and manuscript. MAA was supported by an Australian Commonwealth Research Scholarship; Research was supported by the School of Biological Science, The University of Adelaide.
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Alam, M.A., Kelly, J.M. Proteins interacting with CreA and CreB in the carbon catabolite repression network in Aspergillus nidulans . Curr Genet 63, 669–683 (2017). https://doi.org/10.1007/s00294-016-0667-2
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DOI: https://doi.org/10.1007/s00294-016-0667-2