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Nano-LC-Q-TOF Analysis of Proteome Revealed Germination of Aspergillus flavus Conidia is Accompanied by MAPK Signalling and Cell Wall Modulation

Abstract

Aspergillus flavus is the second most leading cause of aspergillosis. The ability of A. flavus to adapt within the host environment is crtical for its colonization. Onset of germination of conidia is one of the crucial events; thus, in order to gain insight into A. flavus molecular adaptation while germination, protein profile of A. flavus was obtained. Approximately 82 % of conidia showed germination at 7 h; thus, samples were collected followed by protein extraction and subjected to nLC-Q-TOF mass spectrometer. Q-TOF data were analysed using Protein Lynx Global Services (PLGS 2.2.5) software. A total of 416 proteins were identified from UniProt Aspergillus species database. Orthologues of A. flavus was observed in A. fumigatus, A. niger, A. terreus, A. oryzae, etc. Proteins were further analysed in NCBI database, which showed that 27 proteins of A. flavus are not reported in UniProt and NCBI database. Functional characterization of proteins resulted majorly to cell wall synthesis and degradation, metabolisms (carbohydrate and amino acid metabolism), protein synthesis and degradation. Proteins/enzymes associated with aflatoxin biosynthesis were observed. We also observed Dicer-like proteins 1, 2 and autophagy-related proteins 2, 9, 18, 13, 11, 22. Expression of protein/enzymes associated with MAPK signalling pathway suggests their role during the germination process. Overall, the data present a catalogue of proteins/enzymes involved in the germination of A. flavus conidia and could also be applied to other Aspergillus species.

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Acknowledgments

Authors are thankful to Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, for providing facilities and Ph.D. fellowship to ST and RT.

Authors Contributions

ST and JS conceived and designed the experiments. ST performed the experiments. ST, RT and JS analysed the data. GG and JS contributed reagents/materials/analysis tools. ST and JS contributed to writing the manuscript.

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Correspondence to Jata Shankar.

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The authors declare that they have no conflit of interests.

Electronic supplementary material

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Supplementary Table S1 (A)

List of 416 proteins of Aspergillus flavus that are expressed at germinating stage (7 h). (XLSX 131 kb)

Supplementary Table S1 (B)

List of proteins identified in Aspergillus flavus grown on Sabouraud dextrose media for 7 h (germinating conidial stage). Proteins were separated and identified via nL-Q-TOF mass spectroscopy and are clustered according to biological process as defined by Gene Ontology from UniProt database. (XLSX 261 kb)

Supplementary Table S2

Details of selected protein of carbohydrate metabolism involved in various biological, molecular and cellular processes. The data represent the significance in relation to p values and the protein is represented by Gene Ontology IDs obtained from FungiFun 2.2.8 BETA software. (PDF 254 kb)

Supplementary material 4 (DOCX 19 kb)

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Tiwari, S., Thakur, R., Goel, G. et al. Nano-LC-Q-TOF Analysis of Proteome Revealed Germination of Aspergillus flavus Conidia is Accompanied by MAPK Signalling and Cell Wall Modulation. Mycopathologia 181, 769–786 (2016). https://doi.org/10.1007/s11046-016-0056-x

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Keywords

  • Aspergillus flavus
  • Germinating conidia
  • nLC-Q-TOF
  • Proteome
  • Cell wall
  • Anti-Aspergillus targets