Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells

ABSTRACT Xylella fastidiosa (Xf) is a Gram‐negative bacterial plant pathogen responsible for severe diseases in a variety of economically important crops. A critical aspect of its virulence is the production of extracellular vesicles (EVs). In this study, we discovered that DNA‐binding proteins and...

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Main Authors: Alessa Ruf, Patrick Blumenkamp, Christina Ludwig, Anne Lippegaus, Andreas Brachmann, Andreas Klingl, Alexander Goesmann, Karina Brinkrolf, Kai Papenfort, Silke Robatzek
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:Journal of Extracellular Vesicles
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Online Access:https://doi.org/10.1002/jev2.70102
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author Alessa Ruf
Patrick Blumenkamp
Christina Ludwig
Anne Lippegaus
Andreas Brachmann
Andreas Klingl
Alexander Goesmann
Karina Brinkrolf
Kai Papenfort
Silke Robatzek
author_facet Alessa Ruf
Patrick Blumenkamp
Christina Ludwig
Anne Lippegaus
Andreas Brachmann
Andreas Klingl
Alexander Goesmann
Karina Brinkrolf
Kai Papenfort
Silke Robatzek
author_sort Alessa Ruf
collection DOAJ
description ABSTRACT Xylella fastidiosa (Xf) is a Gram‐negative bacterial plant pathogen responsible for severe diseases in a variety of economically important crops. A critical aspect of its virulence is the production of extracellular vesicles (EVs). In this study, we discovered that DNA‐binding proteins and nonribosomal RNA‐binding proteins are abundant in the corona of Xf‐EVs. DNA‐seq revealed enrichment of three genomic islands (GIs) in EVs, which carry molecular signatures indicative of horizontal gene transfer (HGT). The most abundant GI encodes five homologous small RNAs designated sXFs. RNA sequencing revealed a distinct pattern of noncoding RNAs enriched in EVs, including four island‐encoded sXFs. One of the sXF’s stem‐loops contains motifs for binding the RNA chaperone Hfq, which is also abundant in EVs. Predicted target analysis suggests that sXFs play a role in regulation of natural competence in bacteria. Additionally, sXF plant target prediction identifies a coiled‐coil nucleotide‐binding domain leucine‐rich repeat receptor (CNL) immune gene that is downregulated following Xf infection and Xf‐EV treatment. We propose a model where Xf releases nucleic acid carrying EVs with two functions: one to deliver RNA‐related cargo that regulates gene expression in both bacterial and plant cells, and another to deliver DNA‐related cargo for the genetic transfer of genomic islands. We highlight island‐encoded sXFs as potential virulence factors and vesiduction as a mechanism of HGT of sXFs in Xf. Taken together, our data on Xf‐EV cargoes provide a molecular framework for understanding the virulence of Xf.
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spelling doaj-art-6fcb43bc55d147389ff6d86f3eeddfaf2025-08-20T02:48:54ZengWileyJournal of Extracellular Vesicles2001-30782025-06-01146n/an/a10.1002/jev2.70102Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient CellsAlessa Ruf0Patrick Blumenkamp1Christina Ludwig2Anne Lippegaus3Andreas Brachmann4Andreas Klingl5Alexander Goesmann6Karina Brinkrolf7Kai Papenfort8Silke Robatzek9LMU Munich Biocenter Ludwig‐Maximilian‐University of Munich Martinsried GermanyBioinformatics and Systems Biology Justus Liebig University Giessen Giessen GermanyBavarian Center for Biomolecular Mass Spectrometry (BayBioMS) TUM School of Life Sciences Freising GermanyBioinstrumentezentrum Friedrich‐ Schiller University Institute of Microbiology Jena GermanyLMU Munich Biocenter Ludwig‐Maximilian‐University of Munich Martinsried GermanyLMU Munich Biocenter Ludwig‐Maximilian‐University of Munich Martinsried GermanyBioinformatics and Systems Biology Justus Liebig University Giessen Giessen GermanyBioinformatics and Systems Biology Justus Liebig University Giessen Giessen GermanyBioinstrumentezentrum Friedrich‐ Schiller University Institute of Microbiology Jena GermanyLMU Munich Biocenter Ludwig‐Maximilian‐University of Munich Martinsried GermanyABSTRACT Xylella fastidiosa (Xf) is a Gram‐negative bacterial plant pathogen responsible for severe diseases in a variety of economically important crops. A critical aspect of its virulence is the production of extracellular vesicles (EVs). In this study, we discovered that DNA‐binding proteins and nonribosomal RNA‐binding proteins are abundant in the corona of Xf‐EVs. DNA‐seq revealed enrichment of three genomic islands (GIs) in EVs, which carry molecular signatures indicative of horizontal gene transfer (HGT). The most abundant GI encodes five homologous small RNAs designated sXFs. RNA sequencing revealed a distinct pattern of noncoding RNAs enriched in EVs, including four island‐encoded sXFs. One of the sXF’s stem‐loops contains motifs for binding the RNA chaperone Hfq, which is also abundant in EVs. Predicted target analysis suggests that sXFs play a role in regulation of natural competence in bacteria. Additionally, sXF plant target prediction identifies a coiled‐coil nucleotide‐binding domain leucine‐rich repeat receptor (CNL) immune gene that is downregulated following Xf infection and Xf‐EV treatment. We propose a model where Xf releases nucleic acid carrying EVs with two functions: one to deliver RNA‐related cargo that regulates gene expression in both bacterial and plant cells, and another to deliver DNA‐related cargo for the genetic transfer of genomic islands. We highlight island‐encoded sXFs as potential virulence factors and vesiduction as a mechanism of HGT of sXFs in Xf. Taken together, our data on Xf‐EV cargoes provide a molecular framework for understanding the virulence of Xf.https://doi.org/10.1002/jev2.70102EVsgenomic islandsHfqhorizontal gene transfersmall noncoding RNAs
spellingShingle Alessa Ruf
Patrick Blumenkamp
Christina Ludwig
Anne Lippegaus
Andreas Brachmann
Andreas Klingl
Alexander Goesmann
Karina Brinkrolf
Kai Papenfort
Silke Robatzek
Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
Journal of Extracellular Vesicles
EVs
genomic islands
Hfq
horizontal gene transfer
small noncoding RNAs
title Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
title_full Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
title_fullStr Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
title_full_unstemmed Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
title_short Extracellular Vesicles From Xylella fastidiosa Carry sRNAs and Genomic Islands, Suggesting Roles in Recipient Cells
title_sort extracellular vesicles from xylella fastidiosa carry srnas and genomic islands suggesting roles in recipient cells
topic EVs
genomic islands
Hfq
horizontal gene transfer
small noncoding RNAs
url https://doi.org/10.1002/jev2.70102
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