1. Home
  2. Projects Index
  3. Z2

Z2

Overview

In project Z2 a sophisticated proteome analysis platform will be provided based on 2D-gel electrophoresis (2D-GE), MALDI-TOF and LC-MS/MS techniques to specific A and Cprojects.

Programme: Collaborative Research Center / Transregio 124 "Pathogenic fungi and their human host: Networks of Interaction - FungiNet"

SEEK ID: https://funginet.hki-jena.de/projects/35

Public web page: http://www.funginet.de/project-z2.html

Organisms: Aspergillus fumigatus, Candida albicans

INF PALs: No PALs for this Project

Project created: 4th Mar 2016

Related items

Daniela Albrecht-Eckardt

Projects: INF, Z2

Institutions: BioControl Jena GmbH

Peter Großmann programme_administrator: inactive since 2020-06-01

Projects: FungiNet total, INF, A1, A2, A3, A4 (E), A5, A6, B1, B3 (E), B4, B5, C1, C2, C3, C4 (E), C5, C6 (E), Z1, Z2, FungiNet A - Aspergillus projects, FungiNet B - Bioinformatics projects, FungiNet C - Candida projects, B2

Institutions: Leibniz-Institute for Natural Product Research and Infection Biology Hans Knöll Institute (HKI)

https://orcid.org/0000-0001-6106-724X
Franziska Hoffmann

Projects: Z2

Institutions: University Hospital Jena

https://orcid.org/0000-0002-6872-924X
Olaf Kniemeyer project_administrator

Projects: FungiNet total, Z2

Institutions: Leibniz-Institute for Natural Product Research and Infection Biology Hans Knöll Institute (HKI)

https://orcid.org/0000-0002-9493-6402
Thomas Krüger

Projects: FungiNet total, Z2

Institutions: Leibniz-Institute for Natural Product Research and Infection Biology Hans Knöll Institute (HKI)

Gene regulation by hapX
Z2

P Hortschansky, M Eisendle, Q Al-Abdallah, A D Schmidt, S Bergmann, M Thön, O Kniemeyer, B Abt, B Seeber, E R Werner, M Kato, A A Brakhage and H Haas. Interaction of HapX with the CCAAT-binding complex-a novel mechanism of gene regulation by iron. EMBO J. 2007 Jul 11;26(13):3157-68. Epub 2007 Jun 14

View

Snapshots: No snapshots

Farnesol
Z2

Wartenberg D, Vödisch M, Kniemeyer O, Albrecht-Eckardt D, Scherlach K, Winkler R, Weide M, Brakhage AA. Proteome analysis of the farnesol-induced stress response in Aspergillus nidulans-The role of a putative dehydrin. J Proteomics. 2012 May 23.

View

Snapshots: No snapshots

Temperature Shift Proteomic
Z2

D Albrecht, R Guthke, A A Brakhage, O Kniemeyer. Integrative analysis of the heat shock response in Aspergillus fumigatus. BMC Genomics. 2010 Jan 15;11:32.

View

Find corresponding transcriptomic data at ArrayExpress: E-MEXP-332

Snapshots: No snapshots

Secretome AMM/72 h
Z2

Dirk Wartenberg, Katrin Lapp, Ilse D. Jacobsen, Hans-Martin Dahse, Olaf Kniemeyer, Thorsten Heinekamp, Axel A. Brakhage
Leibniz Institute for Natural Product Research and Infection Biology-Hans-Knoell-Institute (HKI), Jena, Germany. Secretome analysis of Aspergillus fumigatus reveals Asp-hemolysin as a major secreted protein. Int J Med Microbiol. 2011, Nov;301(7):602-11

View

Snapshots: No snapshots

Proteomic profiling of serological responses to A.fum antigens (IA)
Z2

Teutschbein J, Simon S, Lother J, Springer J, Hortschansky P, Morton CO, Löffler J, Einsele H, Conneally E, Rogers TR, Guthke R, Brakhage AA, Kniemeyer O. Proteomic profiling of serological responses to Aspergillus fumigatus antigens in patients with invasive aspergillosis. J Proteome Res 2016 Mar 24

View

Snapshots: No snapshots

21 hidden items
deltaHapX
Z2
No description specified

Person responsible: Daniela Albrecht-Eckardt

Snapshots: No snapshots

wt
Z2
No description specified

Person responsible: Daniela Albrecht-Eckardt

Snapshots: No snapshots

Internal Standard
Z2

.

Person responsible: Daniela Albrecht-Eckardt

Snapshots: No snapshots

Farnesol Treatment
Z2

Aspergillus nidulans R21 was grown over night in AMM at 37°C and 200 rpm. The culture was treated with 50 µM farnesol or the same volume of ethanol without farnesol as control. The culture was harvested after 3 hours of further incubation.

Person responsible: Daniela Albrecht-Eckardt

Snapshots: No snapshots

Control
Z2
No description specified

Person responsible: Daniela Albrecht-Eckardt

Snapshots: No snapshots

20 hidden items
48h
Z2
No description specified

Contributor: Daniela Albrecht-Eckardt

Assay type: Experimental Assay Type

Technology type: Technology Type

Snapshots: No snapshots

deltaHapX
Z2
No description specified

Contributor: Daniela Albrecht-Eckardt

Assay type: Experimental Assay Type

Technology type: Technology Type

Snapshots: No snapshots

wt
Z2
No description specified

Contributor: Daniela Albrecht-Eckardt

Assay type: Experimental Assay Type

Technology type: Technology Type

Snapshots: No snapshots

Standard gels
Z2
No description specified

Contributor: Daniela Albrecht-Eckardt

Assay type: Experimental Assay Type

Technology type: Technology Type

Snapshots: No snapshots

Farnesol gels
Z2
No description specified

Contributor: Daniela Albrecht-Eckardt

Assay type: Experimental Assay Type

Technology type: Technology Type

Snapshots: No snapshots

20 hidden items
deltaHapX data
Z2
No description specified

Creator: Daniela Albrecht-Eckardt

Contributor: Daniela Albrecht-Eckardt

Download
wt data
Z2
No description specified

Creator: Daniela Albrecht-Eckardt

Contributor: Daniela Albrecht-Eckardt

Download
Standard data
Z2
No description specified

Creator: Daniela Albrecht-Eckardt

Contributor: Daniela Albrecht-Eckardt

Download
Farnesol data
Z2
No description specified

Creator: Daniela Albrecht-Eckardt

Contributor: Daniela Albrecht-Eckardt

Download
Control data
Z2
No description specified

Creator: Daniela Albrecht-Eckardt

Contributor: Daniela Albrecht-Eckardt

Download
20 hidden items
PAS staining of tissue sections
Z2
No description specified

Creator: Franziska Hoffmann

Contributor: Franziska Hoffmann

Download
H&E staining of tissue sections
Z2
No description specified

Creator: Franziska Hoffmann

Contributor: Franziska Hoffmann

Download
MALDI-Imaging workflow for cryo tissue sections
Z2
No description specified

Creators: Franziska Hoffmann, Daniela Pelzel

Contributor: Franziska Hoffmann

Download
MALDI-Imaging workflow for FFPE tissue sections
Z2
No description specified

Creators: Franziska Hoffmann, Daniela Pelzel

Contributor: Franziska Hoffmann

Download
Formalin Fixation and Paraffin Embedding of tissue samples
Z2
No description specified

Creator: Franziska Hoffmann

Contributor: Franziska Hoffmann

Download
11 hidden items
Proteome Analysis Reveals the Conidial Surface Protein CcpA Essential for Virulence of the Pathogenic Fungus Aspergillus fumigatus.
FungiNet total, Z2

Abstract (Expand)

Aspergillus fumigatus is a common airborne fungal pathogen of humans and a significant source of mortality in immunocompromised individuals. Here, we provide the most extensive cell wall proteome … profiling to date of A. fumigatus resting conidia, the fungal morphotype pertinent to first contact with the host. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identified proteins within the conidial cell wall by hydrogen-fluoride (HF)-pyridine extraction and proteins exposed on the surface using a trypsin-shaving approach. One protein, designated conidial cell wall protein A (CcpA), was identified by both methods and was found to be nearly as abundant as hydrophobic rodlet layer-forming protein RodA. CcpA, an amphiphilic protein, like RodA, peaks in expression during sporulation on resting conidia. Despite high cell wall abundance, the cell surface structure of DeltaccpA resting conidia appeared normal. However, trypsin shaving of DeltaccpA conidia revealed novel surface-exposed proteins not detected on conidia of the wild-type strain. Interestingly, the presence of swollen DeltaccpA conidia led to higher activation of neutrophils and dendritic cells than was seen with wild-type conidia and caused significantly less damage to epithelial cells in vitro In addition, virulence was highly attenuated when cortisone-treated, immunosuppressed mice were infected with DeltaccpA conidia. CcpA-specific memory T cell responses were detectable in healthy human donors naturally exposed to A. fumigatus conidia, suggesting a role for CcpA as a structural protein impacting conidial immunogenicity rather than possessing a protein-intrinsic immunosuppressive effect. Together, these data suggest that CcpA serves as a conidial stealth protein by altering the conidial surface structure to minimize innate immune recognition.IMPORTANCE The mammalian immune system relies on recognition of pathogen surface antigens for targeting and clearance. In the absence of immune evasion strategies, pathogen clearance is rapid. In the case of Aspergillus fumigatus, the successful fungus must avoid phagocytosis in the lung to establish invasive infection. In healthy individuals, fungal spores are cleared by immune cells; however, in immunocompromised patients, clearance mechanisms are impaired. Here, using proteome analyses, we identified CcpA as an important fungal spore protein involved in pathogenesis. A. fumigatus lacking CcpA was more susceptible to immune recognition and prompt eradication and, consequently, exhibited drastically attenuated virulence. In infection studies, CcpA was required for virulence in infected immunocompromised mice, suggesting that it could be used as a possible immunotherapeutic or diagnostic target in the future. In summary, our report adds a protein to the list of those known to be critical to the complex fungal spore surface environment and, more importantly, identifies a protein important for conidial immunogenicity during infection.

Authors: V. Voltersen, M. G. Blango, S. Herrmann, F. Schmidt, Thorsten Heinekamp, M. Strassburger, Thomas Krüger, P. Bacher, Jasmin Lother, Esther Weiß, Kerstin Hünniger, H. Liu, P. Hortschansky, A. Scheffold, Jürgen Löffler, S. Krappmann, S. Nietzsche, Oliver Kurzai, Hermann Einsele, Olaf Kniemeyer, S. G. Filler, U. Reichard, Axel Brakhage

Date Published: No date defined

Journal: MBio

Carbon Catabolite Repression in Filamentous Fungi Is Regulated by Phosphorylation of the Transcription Factor CreA.
FungiNet total, A1, Z2

Abstract (Expand)

Filamentous fungi of the genus Aspergillus are of particular interest for biotechnological applications due to their natural capacity to secrete carbohydrate-active enzymes (CAZy) that target plant … biomass. The presence of easily metabolizable sugars such as glucose, whose concentrations increase during plant biomass hydrolysis, results in the repression of CAZy-encoding genes in a process known as carbon catabolite repression (CCR), which is undesired for the purpose of large-scale enzyme production. To date, the C2H2 transcription factor CreA has been described as the major CC repressor in Aspergillus spp., although little is known about the role of posttranslational modifications in this process. In this work, phosphorylation sites were identified by mass spectrometry on Aspergillus nidulans CreA, and subsequently, the previously identified but uncharacterized site S262, the characterized site S319, and the newly identified sites S268 and T308 were chosen to be mutated to nonphosphorylatable residues before their effect on CCR was investigated. Sites S262, S268, and T308 are important for CreA protein accumulation and cellular localization, DNA binding, and repression of enzyme activities. In agreement with a previous study, site S319 was not important for several here-tested phenotypes but is key for CreA degradation and induction of enzyme activities. All sites were shown to be important for glycogen and trehalose metabolism. This study highlights the importance of CreA phosphorylation sites for the regulation of CCR. These sites are interesting targets for biotechnological strain engineering without the need to delete essential genes, which could result in undesired side effects.IMPORTANCE In filamentous fungi, the transcription factor CreA controls carbohydrate metabolism through the regulation of genes encoding enzymes required for the use of alternative carbon sources. In this work, phosphorylation sites were identified on Aspergillus nidulans CreA, and subsequently, the two newly identified sites S268 and T308, the previously identified but uncharacterized site S262, and the previously characterized site S319 were chosen to be mutated to nonphosphorylatable residues before their effect on CCR was characterized. Sites S262, S268, and T308 are important for CreA protein accumulation and cellular localization, DNA binding, and repression of enzyme activities. In agreement with a previous study, site S319 is not important for several here-tested phenotypes but is key for CreA degradation and induction of enzyme activities. This work characterized novel CreA phosphorylation sites under carbon catabolite-repressing conditions and showed that they are crucial for CreA protein turnover, control of carbohydrate utilization, and biotechnologically relevant enzyme production.

Authors: L. J. de Assis, L. P. Silva, O. Bayram, P. Dowling, Olaf Kniemeyer, Thomas Krüger, Axel Brakhage, Y. Chen, L. Dong, K. Tan, K. H. Wong, L. N. A. Ries, G. H. Goldman

Date Published: 5th Jan 2021

Journal: mBio

Functional surface proteomic profiling reveals the host heat-shock protein A8 as a mediator of Lichtheimia corymbifera recognition by murine alveolar macrophages.
A1, A5, B4, Z2, A6

Abstract (Expand)

Mucormycosis is an emergent, fatal fungal infection of humans and warm-blooded animals caused by species of the order Mucorales. Immune cells of the innate immune system serve as the first line of … defence against inhaled spores. Alveolar macrophages were challenged with the mucoralean fungus Lichtheimia corymbifera and subjected to biotinylation and streptavidin enrichment procedures followed by LC-MS/MS analyses. A total of 28 host proteins enriched for binding to macrophage-L. corymbifera interaction. Among those, the HSP70-family protein Hspa8 was found to be predominantly responsive to living and heat-killed spores of a virulent and an attenuated strain of L. corymbifera. Confocal scanning laser microscopy of infected macrophages revealed colocalization of Hspa8 with phagocytosed spores of L. corymbifera. The amount of detectable Hspa8 was dependent on the multiplicity of infection. Incubation of alveolar macrophages with an anti-Hspa8 antibody prior to infection reduced their capability to phagocytose spores of L. corymbifera. In contrast, anti-Hspa8 antibodies did not abrogate the phagocytosis of Aspergillus fumigatus conidia by macrophages. These results suggest an important contribution of the heat-shock family protein Hspa8 in the recognition of spores of the mucoralean fungus L. corymbifera by host alveolar macrophages and define a potential immunomodulatory therapeutic target.

Authors: M. I. A. Hassan, J. M. Kruse, Thomas Krüger, H. M. Dahse, Z. Cseresnyes, M. G. Blango, Hortense Slevogt, F. Horhold, V. Ast, R. Konig, Marc Thilo Figge, Olaf Kniemeyer, Axel Brakhage, Kerstin Voigt

Date Published: 26th Jun 2020

Journal: Environ Microbiol

Microdissection-An Essential Prerequisite for Spatial Cancer Omics.
Z2

Abstract (Expand)

The problem with cancer tissue is that its intratumoral heterogeneity and its complexity is extremely high as cells possess, depending on their location and function, different mutations, different … mRNA expression and the highest intricacy in the protein pattern. Prior to genomic and proteomic analyses, it is therefore indispensable to identify the exact part of the tissue or even the exact cell. Laser-based microdissection is a tried and tested technique able to produce pure and well-defined cell material for further analysis with proteomic and genomic techniques. It sheds light on the heterogeneity of cancer or other complex diseases and enables the identification of biomarkers. This review aims to raise awareness for the reconsideration of laser-based microdissection and seeks to present current state-of-the-art combinations with omic techniques.

Authors: Ferdinand Von Eggeling, Franziska Hoffmann

Date Published: 25th Jun 2020

Journal: Proteomics

Ahr1 and Tup1 Contribute to the Transcriptional Control of Virulence-Associated Genes in Candida albicans.
A1, C1, C2, C3, C5, Z2

Abstract (Expand)

The capacity of Candida albicans to reversibly change its morphology between yeast and filamentous stages is crucial for its virulence. Formation of hyphae correlates with the upregulation of genes … ALS3 and ECE1, which are involved in pathogenicity processes such as invasion, iron acquisition, and host cell damage. The global repressor Tup1 and its cofactor Nrg1 are considered to be the main antagonists of hyphal development in C. albicans However, our experiments revealed that Tup1, but not Nrg1, was required for full expression of ALS3 and ECE1 In contrast to NRG1, overexpression of TUP1 was found to inhibit neither filamentous growth nor transcription of ALS3 and ECE1 In addition, we identified the transcription factor Ahr1 as being required for full expression of both genes. A hyperactive version of Ahr1 bound directly to the promoters of ALS3 and ECE1 and induced their transcription even in the absence of environmental stimuli. This regulation worked even in the absence of the crucial hyphal growth regulators Cph1 and Efg1 but was dependent on the presence of Tup1. Overall, our results show that Ahr1 and Tup1 are key contributors in the complex regulation of virulence-associated genes in the different C. albicans morphologies.IMPORTANCE Candida albicans is a major human fungal pathogen and the leading cause of systemic Candida infections. In recent years, Als3 and Ece1 were identified as important factors for fungal virulence. Transcription of both corresponding genes is closely associated with hyphal growth. Here, we describe how Tup1, normally a global repressor of gene expression as well as of filamentation, and the transcription factor Ahr1 contribute to full expression of ALS3 and ECE1 in C. albicans hyphae. Both regulators are required for high mRNA amounts of the two genes to ensure functional relevant protein synthesis and localization. These observations identified a new aspect of regulation in the complex transcriptional control of virulence-associated genes in C. albicans.

Authors: S. Ruben, E. Garbe, Selene Mogavero, Daniela Albrecht-Eckardt, D. Hellwig, A. Hader, Thomas Krüger, K. Gerth, Ilse Jacobsen, O. Elshafee, S. Brunke, Kerstin Hünniger, Olaf Kniemeyer, Axel Brakhage, Joachim Morschhäuser, Bernhard Hube, Slavena Vylkova, Oliver Kurzai, R. Martin

Date Published: 28th Apr 2020

Journal: mBio

16 hidden items
7 hidden items
Powered by
 (v.1.9.1)
About INF | FungiNet | Funding and Programmes | Credits | Imprint
Copyright © 2008 - 2019 The University of Manchester and HITS gGmbH