Polysome profiling reveals broad translatome remodeling during endoplasmic reticulum (ER) stress in the pathogenic fungus Aspergillus fumigatus

BMC Genomics

David S Askew¹  Long Jason Lu²  William C Nierman⁴  Liliana Losada⁴  Zhaowei Ren²  Karthik Krishnan³ 

[1]Department of Pathology, University of Cincinnati, PO Box 670529, Cincinnati, OH 45267-0529, USA

[2]Division of Biomedical Informatics, Cincinnati Children’s Hospital Research Foundation, Cincinnati, OH 45229, USA

[3]Department of Pathology & Laboratory Medicine, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0529, USA

[4]The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA

关键词: Yvc1;    Polysome profiling;    Translational regulation;    ER stress;    Unfolded protein response;    UPR;    Aspergillus fumigatus;   

Others  :  1217837
DOI  :  10.1186/1471-2164-15-159

 received in 2013-10-17, accepted in 2014-02-17,  发布年份 2014

【 摘 要 】

Background

The unfolded protein response (UPR) is a network of intracellular signaling pathways that supports the ability of the secretory pathway to maintain a balance between the load of proteins entering the endoplasmic reticulum (ER) and the protein folding capacity of the ER lumen. Current evidence indicates that several pathogenic fungi rely heavily on this pathway for virulence, but there is limited understanding of the mechanisms involved. The best known functional output of the UPR is transcriptional upregulation of mRNAs involved in ER homeostasis. However, this does not take into account mechanisms of translational regulation that involve differential loading of ribosomes onto mRNAs. In this study, a global analysis of transcript-specific translational regulation was performed in the pathogenic mold Aspergillus fumigatus to determine the nature and scope of the translational response to ER stress.

Results

ER stress was induced by treating the fungus with dithiothreitol, tunicamycin, or a thermal up-shift. The mRNAs were then fractionated on the basis of ribosome occupancy into an under-translated pool (U) and a well-translated pool (W). The mRNAs were used to interrogate microarrays and the ratio of the hybridization signal (W/U) was used as an indicator of the relative translational efficiency of a mRNA under each condition. The largest category of translationally upregulated mRNAs during ER stress encoded proteins involved in translation. Components of the ergosterol and GPI anchor biosynthetic pathways also showed increased polysome association, suggesting an important role for translational regulation in membrane and cell wall homeostasis. ER stress induced limited remodeling of the secretory pathway translatome. However, a select group of transcription factors was translationally upregulated, providing a link to subsequent modification of the transcriptome. Finally, we provide evidence that one component of the ER stress translatome is a novel mRNA isoform from the yvc1 gene that is induced by ER stress in a UPR-dependent manner.

Conclusions

Together, these findings define a core set of mRNAs subject to translational control during the adaptive response to acute ER stress in A. fumigatus and reveal a remarkable breadth of functions that are needed to resolve ER stress in this organism.

【 授权许可】

   
2014 Krishnan et al.; licensee BioMed Central Ltd.

【 图 表 】

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