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Ib101206 - proteome data analysis

ABSTRACT: P2 is a spontaneous mutant selected from P1, an entomopathogenic fungus Beauveria bassiana that hyper-produces extracellular protease(s), compared to P1. The major expressed serine protease, called SBP, is shown to be up-regulated at the transcriptional level in the P2 mutant strain. The insecticidal potential of P1 and P2 strains was evaluated against Tuta absoluta larvae under laboratory conditions. Both strains are effective but P2 showed stronger effect than P1. Median lethal concentration (LC50) of P2 is 26 fold lower than that of P1. Median lethal times (LT50) of P1 and P2 are 4.38 and 0.84 days using 105 conidia ml-1. Enzymatic assays analysis showed that extracellular enzymes are differently expressed by the two strains, especially proteases and chitinases. Two-dimensional gel electrophoresis and mass spectrometry analysis confirmed the overexpression by P2 strain of protease and chitinase and revealed further the over-expression of several protease isoforms. Here we show a direct link between fungal enzymatic profile and insect’s pathogenesis, giving a molecular explanation for the insecticidal potential of B. bassiana fungus.

INSTRUMENT(S): LCQ Classic

ORGANISM(S): Beauveria Bassiana (white Muscardine Disease Fungus) (tritirachium Shiotae)

SUBMITTER: Dupuy Jean-William

LAB HEAD: Ali gargouri

PROVIDER: PXD003287 | Pride | 2018-03-19

REPOSITORIES: Pride

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Project description:Candida albicans is an opportunistic pathogen and a commensal fungus of the mucosa from many healthy patients. Invasive candidiasis (IC) has high levels of morbidity and mortality that are attributed to some extent to the difficulty in its early diagnosis. Diverse and immunogenic proteins with roles such as virulence and immune evasion are secreted by this pathogen. Immunoproteomics is an important technique for the assessment of antibody reactivities to immunogenic proteins and our group previously developed an algorithm to predict the clinical outcome in IC patients. With the ultimate purpose of discovering new biomarkers of IC, immunoproteomics was applied for the analysis of the serological response to hyphal C. albicans extracellular proteins. Gel-free proteomic analysis was performed by LC-MS/MS for the detection of the secreted proteins and 301 proteins were identified. As expected, gene ontology analysis in cellular component showed enrichment in proteins from extracellular region, cell wall, cell surface, cell periphery and intracellular region. Importantly, 46.5% of the intracellular proteins identified were also annotated in extracellular region. Furthermore, from the extracellular proteins identified here, 44 proteins were previously found to be immunogenic against human sera. To carry out immunoproteomic analysis, secreted proteins were separated by two-dimensional gel electrophoresis, electroblotted onto nitrocellulose membranes, and tested by Western blotting with three distinct pools of human sera: P1 (12 IC patients), P2 (11 patients with Candida infection associated with catheters) and P3 (11 patients without IC). A higher reactivity pattern was observed in P1 and P2 in comparison with P3. So far, Glx3 was immunogenic to P1 and P2 but not to P3. Eno1, Pgk1 and Pra1 were found to be more immunoreactive to both P1 and P2 than to the P3. This approach will contribute to a better and faster diagnosis of this life-threatening fungal infection.

			Action	DRS
	UP_2015_09_Beauvaria_bassiana.fasta	Fasta
	ib101206_01.RAW	Raw
	ib101206_01.msf	Msf
	ib101206_02.RAW	Raw
	ib101206_02.msf	Msf

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Ib101206 - proteome data analysis

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