Dataset.

Dynamics of interaction and internalisation of the antifungal protein PeAfpA into Penicillium digitatum morphotypes [DATASET]

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/370423
Digital.CSIC. Repositorio Institucional del CSIC
  • Giner Llorca, Moisés
  • Ropero Pérez, Carolina
  • Garrigues, Sandra
  • Thomson, Darren D.
  • Bignell, Elaine M.
  • Manzanares, Paloma
  • Marcos López, José Francisco
Raw data of manuscript "Dynamics of interaction and internalisation of the antifungal protein PeAfpA into Penicillium digitatum morphotypes", This dataset includes the raw data from the manuscript “Dynamics of interaction and internalisation of the antifungal protein PeAfpA into Penicillium digitatum morphotypes”. This data comprises Excel sheets used to generate all the plots and graphs of the article (.xlsx files), 16-bit, 8-bit and RGB confocal microscopy images (.tif files), statistical analyses (.pzfx and .txt files) and “Readme” files (.txt files) to specificy experimental conditions for data collection and software-specific requirements to visualise each file., This work was supported by grant PID2021-125858OB-I00, and the Severo Ochoa Excellence Program CEX2021-001189-S, funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF, a way of making Europe”. M.G.L. and C.R.P. were recipients of predoctoral grants FPU19/02066 and ACIF/2021/364 funded by MCIN/AEI/10.13039/501100011033 and by ESF-Investing in Your future and by ”Conselleria d’Educació (Generalitat Valenciana, Comunitat Valenciana, Spain), respectively. M.G.L. international exchange was supported by EST23/00233 grant (funded by MCIN/AEI/10.13039/ 501100011033 and by ESF-Investing in Your future) and SEG10314 EMBO Scientific Exchange Grant. S.G. holds a Juan de la Cierva Incorporación grant (IJC2020-042749-I) from the Spanish Ministerio de Ciencia e Innovación, funded by the European Union, NextGenerationEU PRTR. Work undertaken with EMB and DDT was funded by the Medical Research Council Centre for Medical Mycology at the University of Exeter (MR/N006364/2 and MR/V033417/1), that forms part of the National Institute for Health and Care Research (NIHR) Exeter Biomedical Research Centre (BRC), and the BBSRC project grant BB/V017004/1 to EMB., dor (dc.descripction.sponsorship): With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2021-001189-S), Figure 1_data: Antifungal activity of the antifungal proteins PeAfpA and PdAfpB against Penicillium digitatum conidia. -- Figure 2_data ROS production in Penicilliumn digitatum germlings, induced by the treatment with antifungal proteins (PeAfpA or PdAfpB).-- Figure 3_data Evaluation of the interaction of PeAfpA and PdAfpB with P. digitaum conidia. Panels show confocal microscopy images of P. digitatum conidia after 15 min or 90 min of treatment with 8 µg/mL PeAfpA (A, B), or after 90 min of treatment with 8 µg/mL PdAfpB (C).-- Figure 4_data Comparison of the PeAfpA interaction with conidia of P. digitatum wild-type strain CECT 20796 (WT) and the KO mutant strains ΔpksP and Δarp2. -- Figure 5_data BODIPY-PeAfpA internalisation dynamics in P. digitatum hyphae in low-dose conditions (2 µg/mL).-- Figure 6_data PeAfpA internalises through hyphal tips and provokes vacuolisation and hyphal burst. Raw images used for each panel are provided.-- Figure 7_data: PeAfpA co-localises with CFW in the fungal cell wall. Raw images used for each panel are provided.-- Figure 8_data: PeAfpA internalisation in fungal hyphae is energy-dependent but not an endocytic process. Raw images used for each panel are provided.-- Supplementary Figure 1_data: Antifungal activity of BODIPY-labelled and -unlabelled antifungal proteins (PeAfpA and PdAfpB) against Penicillium digitatum conidia. Raw OD600 measurements are provided in two separated Excel files. -- Supplementary Figure 2_data: Raw data for the amino acid alignment of the Aspergillus fumigatus RodA protein and the orthologous protein from Penicillium digitatum. Raw pictures for panel B are provided.-- Supplementary Figure 3_data: Raw OD measurements for the dose response curves of P. digitatum CECT 20796 and deletion mutant strains (ΔpksP and Δarp2) treated with PeAfpA and PdAfpB. -- Supplementary Figure 4_data: Evaluation of the colocalization of PeAfpA and CFW in P. digitatum hyphae. Rae images for each panel are provided, as well as raw pixel intensity data.-- Supplementary Figure 5_data: Raw images from insets shown in Figure 5., Peer reviewed
 

DOI: http://hdl.handle.net/10261/370423, https://doi.org/10.20350/digitalCSIC/16632
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/370423

HANDLE: http://hdl.handle.net/10261/370423, https://doi.org/10.20350/digitalCSIC/16632
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/370423
 
Ver en: http://hdl.handle.net/10261/370423, https://doi.org/10.20350/digitalCSIC/16632
Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/370423

Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/374724
Artículo científico (article). 2024

DYNAMICS OF INTERACTION AND INTERNALISATION OF THE ANTIFUNGAL PROTEIN PEAFPA INTO PENICILLIUM DIGITATUM MORPHOTYPES

Digital.CSIC. Repositorio Institucional del CSIC
  • Giner Llorca, Moisés
  • Ropero Pérez, Carolina
  • Garrigues, Sandra
  • Thomson, Darren D.
  • Bignell, Elaine M.
  • Manzanares, Paloma
  • Marcos López, José Francisco
Antifungal proteins (AFPs) as the highly active PeAfpA from Penicillium expansum or PdAfpB from Penicillium digitatum exert promising antifungal activity, but their mode of action is not fully understood. We characterised the interaction of PeAfpA against P. digitatum, comparing it to the less active PdAfpB. Despite similar effect on conidia germination, PeAfpA did not induce a burst of reactive oxygen species as PdAfpB. Live-cell fluorescence microscopy revealed complex dynamics of interaction and internalisation of both proteins with distinct P. digitatum morphotypes (quiescent conidia, swollen conidia, germlings and hyphae). Labelled PeAfpA co-localised at the cell wall of quiescent conidia, where its localisation was punctate and not uniformly distributed. This pattern changed during germination to a uniform distribution with increased intensity. Conidia from mutants of genes involved in melanin biosynthesis (pksP/alb1 or arp2) showed an altered distribution of PeAfpA but later mimicked the wild type trend of changes during germination. In swollen conidia and germlings, PeAfpA remained attached to the cell wall. In hyphae, PeAfpA was internalised through the growing hyphal tip after binding to the cell wall, in a non-endocytic but energy-dependent process that caused vacuolisation, which preceded cell death. These results may help the development of biofungicides based on AFPs., This work was supported by grant PID2021-125858OB-I00, and the Severo Ochoa Excellence Program CEX2021-001189-S, funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF, a way of making Europe”. M.G.L. and C.R.P. were recipients of predoctoral grants FPU19/02066 and ACIF/2021/364 funded by MCIN/AEI/10.13039/501100011033 and by ESF-Investing in Your future and by “Conselleria d'Educació” (Generalitat Valenciana, Comunitat Valenciana, Spain), respectively. M.G.L. international exchange was supported by EST23/00233 grant (funded by MCIN/AEI/10.13039/ 501100011033 and by ESF-Investing in Your future) and SEG10314 EMBO Scientific Exchange Grant. S.G. holds a Juan de la Cierva Incorporación grant (IJC2020-042749-I) from the Spanish Ministerio de Ciencia e Innovación, funded by the European Union, NextGenerationEU PRTR. Work undertaken with EMB and DDT was funded by the Medical Research Council Centre for Medical Mycology at the University of Exeter (MR/N006364/2 and MR/V033417/1), that forms part of the National Institute for Health and Care Research (NIHR) Exeter Biomedical Research Centre (BRC), and the BBSRC project grant BB/V017004/1 to EMB., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2021-001189-S), Peer reviewed




Digital.CSIC. Repositorio Institucional del CSIC
oai:digital.csic.es:10261/370423
Dataset. 2024

DYNAMICS OF INTERACTION AND INTERNALISATION OF THE ANTIFUNGAL PROTEIN PEAFPA INTO PENICILLIUM DIGITATUM MORPHOTYPES [DATASET]

Digital.CSIC. Repositorio Institucional del CSIC
  • Giner Llorca, Moisés
  • Ropero Pérez, Carolina
  • Garrigues, Sandra
  • Thomson, Darren D.
  • Bignell, Elaine M.
  • Manzanares, Paloma
  • Marcos López, José Francisco
Raw data of manuscript "Dynamics of interaction and internalisation of the antifungal protein PeAfpA into Penicillium digitatum morphotypes", This dataset includes the raw data from the manuscript “Dynamics of interaction and internalisation of the antifungal protein PeAfpA into Penicillium digitatum morphotypes”. This data comprises Excel sheets used to generate all the plots and graphs of the article (.xlsx files), 16-bit, 8-bit and RGB confocal microscopy images (.tif files), statistical analyses (.pzfx and .txt files) and “Readme” files (.txt files) to specificy experimental conditions for data collection and software-specific requirements to visualise each file., This work was supported by grant PID2021-125858OB-I00, and the Severo Ochoa Excellence Program CEX2021-001189-S, funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF, a way of making Europe”. M.G.L. and C.R.P. were recipients of predoctoral grants FPU19/02066 and ACIF/2021/364 funded by MCIN/AEI/10.13039/501100011033 and by ESF-Investing in Your future and by ”Conselleria d’Educació (Generalitat Valenciana, Comunitat Valenciana, Spain), respectively. M.G.L. international exchange was supported by EST23/00233 grant (funded by MCIN/AEI/10.13039/ 501100011033 and by ESF-Investing in Your future) and SEG10314 EMBO Scientific Exchange Grant. S.G. holds a Juan de la Cierva Incorporación grant (IJC2020-042749-I) from the Spanish Ministerio de Ciencia e Innovación, funded by the European Union, NextGenerationEU PRTR. Work undertaken with EMB and DDT was funded by the Medical Research Council Centre for Medical Mycology at the University of Exeter (MR/N006364/2 and MR/V033417/1), that forms part of the National Institute for Health and Care Research (NIHR) Exeter Biomedical Research Centre (BRC), and the BBSRC project grant BB/V017004/1 to EMB., dor (dc.descripction.sponsorship): With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2021-001189-S), Figure 1_data: Antifungal activity of the antifungal proteins PeAfpA and PdAfpB against Penicillium digitatum conidia. -- Figure 2_data ROS production in Penicilliumn digitatum germlings, induced by the treatment with antifungal proteins (PeAfpA or PdAfpB).-- Figure 3_data Evaluation of the interaction of PeAfpA and PdAfpB with P. digitaum conidia. Panels show confocal microscopy images of P. digitatum conidia after 15 min or 90 min of treatment with 8 µg/mL PeAfpA (A, B), or after 90 min of treatment with 8 µg/mL PdAfpB (C).-- Figure 4_data Comparison of the PeAfpA interaction with conidia of P. digitatum wild-type strain CECT 20796 (WT) and the KO mutant strains ΔpksP and Δarp2. -- Figure 5_data BODIPY-PeAfpA internalisation dynamics in P. digitatum hyphae in low-dose conditions (2 µg/mL).-- Figure 6_data PeAfpA internalises through hyphal tips and provokes vacuolisation and hyphal burst. Raw images used for each panel are provided.-- Figure 7_data: PeAfpA co-localises with CFW in the fungal cell wall. Raw images used for each panel are provided.-- Figure 8_data: PeAfpA internalisation in fungal hyphae is energy-dependent but not an endocytic process. Raw images used for each panel are provided.-- Supplementary Figure 1_data: Antifungal activity of BODIPY-labelled and -unlabelled antifungal proteins (PeAfpA and PdAfpB) against Penicillium digitatum conidia. Raw OD600 measurements are provided in two separated Excel files. -- Supplementary Figure 2_data: Raw data for the amino acid alignment of the Aspergillus fumigatus RodA protein and the orthologous protein from Penicillium digitatum. Raw pictures for panel B are provided.-- Supplementary Figure 3_data: Raw OD measurements for the dose response curves of P. digitatum CECT 20796 and deletion mutant strains (ΔpksP and Δarp2) treated with PeAfpA and PdAfpB. -- Supplementary Figure 4_data: Evaluation of the colocalization of PeAfpA and CFW in P. digitatum hyphae. Rae images for each panel are provided, as well as raw pixel intensity data.-- Supplementary Figure 5_data: Raw images from insets shown in Figure 5., Peer reviewed





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