27 results on '"Chane-Woon-Ming B"'
Search Results
2. P0676 : Clearance of persistent hepatitis c virus infection using a monoclonal antibody specific for tight junction protein claudin-1
- Author
-
Mailly, L., primary, Xiao, F., additional, Lupberger, J., additional, Wilson, G.K., additional, Aubert, P., additional, Duong, F.H., additional, Calabrese, D., additional, Leboeuf, C., additional, Fofana, I., additional, Thumann, C., additional, Bandiera, S., additional, Lütgehetmann, M., additional, Volz, T., additional, Davis, C., additional, Harris, H.J., additional, Mee, C., additional, Girardi, E., additional, Chane-Woon-Ming, B., additional, Ericsson, M., additional, Fletcher, N., additional, Bartenschlager, R., additional, Pessaux, P., additional, Vercauteren, K., additional, Meuleman, P., additional, Villa, P., additional, Kaderali, L., additional, Pfeffer, S., additional, Heim, M.H., additional, Neunlist, M., additional, Zeisel, M.B., additional, Dandri, M., additional, McKeating, J.A., additional, Robinet, E., additional, and Baumert, T.F., additional
- Published
- 2015
- Full Text
- View/download PDF
3. Experimental Evolution of a Plant Pathogen into a Legume Symbiont
- Author
-
Walker, GC, Marchetti, M, Capela, D, Glew, M, Cruveiller, S, Chane-Woon-Ming, B, Gris, C, Timmers, T, Poinsot, V, Gilbert, LB, Heeb, P, Medigue, C, Batut, J, Masson-Boivin, C, Walker, GC, Marchetti, M, Capela, D, Glew, M, Cruveiller, S, Chane-Woon-Ming, B, Gris, C, Timmers, T, Poinsot, V, Gilbert, LB, Heeb, P, Medigue, C, Batut, J, and Masson-Boivin, C
- Abstract
Rhizobia are phylogenetically disparate alpha- and beta-proteobacteria that have achieved the environmentally essential function of fixing atmospheric nitrogen in symbiosis with legumes. Ample evidence indicates that horizontal transfer of symbiotic plasmids/islands has played a crucial role in rhizobia evolution. However, adaptive mechanisms that allow the recipient genomes to express symbiotic traits are unknown. Here, we report on the experimental evolution of a pathogenic Ralstonia solanacearum chimera carrying the symbiotic plasmid of the rhizobium Cupriavidus taiwanensis into Mimosa nodulating and infecting symbionts. Two types of adaptive mutations in the hrpG-controlled virulence pathway of R. solanacearum were identified that are crucial for the transition from pathogenicity towards mutualism. Inactivation of the hrcV structural gene of the type III secretion system allowed nodulation and early infection to take place, whereas inactivation of the master virulence regulator hrpG allowed intracellular infection of nodule cells. Our findings predict that natural selection of adaptive changes in the legume environment following horizontal transfer has been a major driving force in rhizobia evolution and diversification and show the potential of experimental evolution to decipher the mechanisms leading to symbiosis.
- Published
- 2010
4. Correction: Molecular and Evolutionary Bases of Within-Patient Genotypic and Phenotypic Diversity in Escherichia coli Extraintestinal Infections
- Author
-
Levert M, Zamfir O, Clermont O, Bouvet O, Lespinats S, Mc, Hipeaux, Branger C, Picard B, Saint-Ruf C, Norel F, Balliau T, Michel Zivy, Le Nagard H, Cruvellier S, Chane-Woon-Ming B, Nilsson S, Gudelj I, Phan K, Ferenci T, and Tenaillon O
5. The helicase domain of human Dicer prevents RNAi-independent activation of antiviral and inflammatory pathways.
- Author
-
Baldaccini M, Gaucherand L, Chane-Woon-Ming B, Messmer M, Gucciardi F, and Pfeffer S
- Subjects
- Animals, Humans, RNA Interference, Interferon Type I, NF-kappa B genetics, DEAD-box RNA Helicases chemistry, DEAD-box RNA Helicases genetics, DEAD-box RNA Helicases metabolism, Ribonuclease III chemistry, Ribonuclease III genetics, Ribonuclease III metabolism, RNA Virus Infections enzymology
- Abstract
In mammalian somatic cells, the relative contribution of RNAi and the type I interferon response during viral infection is unclear. The apparent inefficiency of antiviral RNAi might be due to self-limiting properties and mitigating co-factors of the key enzyme Dicer. In particular, the helicase domain of human Dicer appears to be an important restriction factor of its activity. Here, we study the involvement of several helicase-truncated mutants of human Dicer in the antiviral response. All deletion mutants display a PKR-dependent antiviral phenotype against certain viruses, and one of them, Dicer N1, acts in a completely RNAi-independent manner. Transcriptomic analyses show that many genes from the interferon and inflammatory response pathways are upregulated in Dicer N1 expressing cells. We show that some of these genes are controlled by NF-kB and that blocking this pathway abrogates the antiviral phenotype of Dicer N1. Our findings highlight the crosstalk between Dicer, PKR, and the NF-kB pathway, and suggest that human Dicer may have repurposed its helicase domain to prevent basal activation of antiviral and inflammatory pathways., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
6. Ribosome Profiling Methods Adapted to the Study of RNA-Dependent Translation Regulation in Staphylococcus aureus.
- Author
-
Kohl MP, Chane-Woon-Ming B, Bahena-Ceron R, Jaramillo-Ponce J, Antoine L, Herrgott L, Romby P, and Marzi S
- Subjects
- Animals, Humans, Gene Expression Regulation, RNA, Ribosomal genetics, RNA, Messenger genetics, Gene Expression Regulation, Bacterial, Staphylococcus aureus metabolism, Ribosome Profiling
- Abstract
Noncoding RNAs, including regulatory RNAs (sRNAs), are instrumental in regulating gene expression in pathogenic bacteria, allowing them to adapt to various stresses encountered in their host environments. Staphylococcus aureus is a well-studied model for RNA-mediated regulation of virulence and pathogenicity, with sRNAs playing significant roles in shaping S. aureus interactions with human and animal hosts. By modulating the translation and/or stability of target mRNAs, sRNAs regulate the synthesis of virulence factors and regulatory proteins required for pathogenesis. Moreover, perturbation of the levels of RNA modifications in two other classes of noncoding RNAs, rRNAs, and tRNAs, has been proposed to contribute to stress adaptation. However, the study of how these various factors affect translation regulation has often been restricted to specific genes, using in vivo reporters and/or in vitro translation systems. Genome-wide sequencing approaches offer novel perspectives for studying RNA-dependent regulation. In particular, ribosome profiling methods provide a powerful resource for characterizing the overall landscape of translational regulation, contributing to a better understanding of S. aureus physiopathology. Here, we describe protocols that we have adapted to perform ribosome profiling in S. aureus., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2024
- Full Text
- View/download PDF
7. Proteomics-based determination of double-stranded RNA interactome reveals known and new factors involved in Sindbis virus infection.
- Author
-
Girardi E, Messmer M, Lopez P, Fender A, Chicher J, Chane-Woon-Ming B, Hammann P, and Pfeffer S
- Subjects
- Humans, Proteomics, Sindbis Virus genetics, Sindbis Virus metabolism, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, RNA, Viral genetics, RNA, Viral metabolism, Virus Replication genetics, RNA, Double-Stranded genetics, RNA Viruses genetics
- Abstract
Viruses are obligate intracellular parasites, which depend on the host cellular machineries to replicate their genome and complete their infectious cycle. Long double-stranded (ds)RNA is a common viral by-product originating during RNA virus replication and is universally sensed as a danger signal to trigger the antiviral response. As a result, viruses hide dsRNA intermediates into viral replication factories and have evolved strategies to hijack cellular proteins for their benefit. The characterization of the host factors associated with viral dsRNA and involved in viral replication remains a major challenge to develop new antiviral drugs against RNA viruses. Here, we performed anti-dsRNA immunoprecipitation followed by mass spectrometry analysis to fully characterize the dsRNA interactome in Sindbis virus (SINV) infected human cells. Among the identified proteins, we characterized SFPQ (splicing factor, proline-glutamine rich) as a new dsRNA-associated proviral factor upon SINV infection. We showed that SFPQ depletion reduces SINV infection in human HCT116 and SK-N-BE(2) cells, suggesting that SFPQ enhances viral production. We demonstrated that the cytoplasmic fraction of SFPQ partially colocalizes with dsRNA upon SINV infection. In agreement, we proved by RNA-IP that SFPQ can bind dsRNA and viral RNA. Furthermore, we showed that overexpression of a wild-type, but not an RNA binding mutant SFPQ, increased viral infection, suggesting that RNA binding is essential for its positive effect on the virus. Overall, this study provides the community with a compendium of dsRNA-associated factors during viral infection and identifies SFPQ as a new proviral dsRNA binding protein., (© 2023 Girardi et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
- Published
- 2023
- Full Text
- View/download PDF
8. Human DICER helicase domain recruits PKR and modulates its antiviral activity.
- Author
-
Montavon TC, Baldaccini M, Lefèvre M, Girardi E, Chane-Woon-Ming B, Messmer M, Hammann P, Chicher J, and Pfeffer S
- Subjects
- Alphavirus Infections metabolism, Alphavirus Infections pathology, DEAD-box RNA Helicases genetics, HEK293 Cells, Humans, Interferon Type I pharmacology, Ribonuclease III genetics, eIF-2 Kinase genetics, Alphavirus Infections drug therapy, Antiviral Agents pharmacology, DEAD-box RNA Helicases metabolism, Protein Interaction Domains and Motifs drug effects, Ribonuclease III metabolism, Semliki forest virus drug effects, Virus Replication, eIF-2 Kinase metabolism
- Abstract
The antiviral innate immune response mainly involves type I interferon (IFN) in mammalian cells. The contribution of the RNA silencing machinery remains to be established, but several recent studies indicate that the ribonuclease DICER can generate viral siRNAs in specific conditions. It has also been proposed that type I IFN and RNA silencing could be mutually exclusive antiviral responses. In order to decipher the implication of DICER during infection of human cells with alphaviruses such as the Sindbis virus and Semliki forest virus, we determined its interactome by proteomics analysis. We show that DICER specifically interacts with several double-stranded RNA binding proteins and RNA helicases during viral infection. In particular, proteins such as DHX9, ADAR-1 and the protein kinase RNA-activated (PKR) are enriched with DICER in virus-infected cells. We demonstrate that the helicase domain of DICER is essential for this interaction and that its deletion confers antiviral properties to this protein in an RNAi-independent, PKR-dependent, manner., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
- Full Text
- View/download PDF
9. Comparative patterns of modified nucleotides in individual tRNA species from a mesophilic and two thermophilic archaea.
- Author
-
Wolff P, Villette C, Zumsteg J, Heintz D, Antoine L, Chane-Woon-Ming B, Droogmans L, Grosjean H, and Westhof E
- Subjects
- Base Sequence, Nucleic Acid Conformation, RNA, Archaeal chemistry, RNA, Archaeal genetics, Methanococcus genetics, Nucleotides chemistry, Pyrococcus furiosus genetics, RNA, Transfer chemistry, RNA, Transfer genetics, Sulfolobus acidocaldarius genetics
- Abstract
To improve and complete our knowledge of archaeal tRNA modification patterns, we have identified and compared the modification pattern (type and location) in tRNAs of three very different archaeal species, Methanococcus maripaludis (a mesophilic methanogen), Pyrococcus furiosus (a hyperthermophile thermococcale), and Sulfolobus acidocaldarius (an acidophilic thermophilic sulfolobale). Most abundant isoacceptor tRNAs (79 in total) for each of the 20 amino acids were isolated by two-dimensional gel electrophoresis followed by in-gel RNase digestions. The resulting oligonucleotide fragments were separated by nanoLC and their nucleotide content analyzed by mass spectrometry (MS/MS). Analysis of total modified nucleosides obtained from complete digestion of bulk tRNAs was also performed. Distinct base- and/or ribose-methylations, cytidine acetylations, and thiolated pyrimidines were identified, some at new positions in tRNAs. Novel, some tentatively identified, modifications were also found. The least diversified modification landscape is observed in the mesophilic Methanococcus maripaludis and the most complex one in Sulfolobus acidocaldarius Notable observations are the frequent occurrence of ac
4 C nucleotides in thermophilic archaeal tRNAs, the presence of m7 G at positions 1 and 10 in Pyrococcus furiosus tRNAs, and the use of wyosine derivatives at position 37 of tRNAs, especially those decoding U1- and C1-starting codons. These results complete those already obtained by others with sets of archaeal tRNAs from Methanocaldococcus jannaschii and Haloferax volcanii ., (© 2020 Wolff et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)- Published
- 2020
- Full Text
- View/download PDF
10. High-Throughput Fluorescence-Based Screen Identifies the Neuronal MicroRNA miR-124 as a Positive Regulator of Alphavirus Infection.
- Author
-
López P, Girardi E, Mounce BC, Weiss A, Chane-Woon-Ming B, Messmer M, Kaukinen P, Kopp A, Bortolamiol-Becet D, Fendri A, Vignuzzi M, Brino L, and Pfeffer S
- Subjects
- Alphavirus metabolism, Alphavirus Infections diagnosis, Cell Line, Chikungunya Fever genetics, Chikungunya virus genetics, Fluorescence, High-Throughput Screening Assays methods, Host-Pathogen Interactions, Humans, MicroRNAs metabolism, Neurons metabolism, RNA, Viral metabolism, Sindbis Virus genetics, Virus Replication, Alphavirus genetics, Alphavirus Infections genetics, MicroRNAs genetics
- Abstract
MicroRNAs (miRNAs) are small regulatory RNAs which act by modulating the expression of target genes. In addition to their role in maintaining essential physiological functions in the cell, miRNAs can also regulate viral infections. They can do so directly by targeting RNAs of viral origin or indirectly by targeting host mRNAs, and this can result in a positive or negative outcome for the virus. Here, we performed a fluorescence-based miRNA genome-wide screen in order to identify cellular miRNAs involved in the regulation of arbovirus infection in human cells. We identified 16 miRNAs showing a positive effect on Sindbis virus (SINV) expressing green fluorescent protein (GFP), among which were a number of neuron-specific ones such as miR-124. We confirmed that overexpression of miR-124 increases both SINV structural protein translation and viral production and that this effect is mediated by its seed sequence. We further demonstrated that the SINV genome possesses a binding site for miR-124. Both inhibition of miR-124 and silent mutations to disrupt this binding site in the viral RNA abolished positive regulation. We also proved that miR-124 inhibition reduces SINV infection in human differentiated neuronal cells. Finally, we showed that the proviral effect of miR-124 is conserved in other alphaviruses, as its inhibition reduces chikungunya virus (CHIKV) production in human cells. Altogether, our work expands the panel of positive regulation of the viral cycle by direct binding of host miRNAs to the viral RNA and provides new insights into the role of cellular miRNAs as regulators of alphavirus infection. IMPORTANCE Arthropod-borne (arbo) viruses are part of a class of pathogens that are transmitted to their final hosts by insects. Because of climate change, the habitat of some of these insects, such as mosquitoes, is shifting, thereby facilitating the emergence of viral epidemics. Among the pathologies associated with arbovirus infection, neurological diseases such as meningitis and encephalitis represent a significant health burden. Using a genome-wide miRNA screen, we identified neuronal miR-124 as a positive regulator of the Sindbis and chikungunya alphaviruses. We also showed that this effect was in part direct, thereby opening novel avenues to treat alphavirus infections., (Copyright © 2020 American Society for Microbiology.)
- Published
- 2020
- Full Text
- View/download PDF
11. Functional microRNA screen uncovers O-linked N-acetylglucosamine transferase as a host factor modulating hepatitis C virus morphogenesis and infectivity.
- Author
-
Herzog K, Bandiera S, Pernot S, Fauvelle C, Jühling F, Weiss A, Bull A, Durand SC, Chane-Woon-Ming B, Pfeffer S, Mercey M, Lerat H, Meunier JC, Raffelsberger W, Brino L, Baumert TF, and Zeisel MB
- Subjects
- Gene Expression Regulation physiology, Gene Knockdown Techniques methods, Genome-Wide Association Study methods, Hepacivirus physiology, Hepatitis C, Chronic virology, Hepatocytes virology, Host-Pathogen Interactions genetics, Humans, Life Cycle Stages genetics, MicroRNAs genetics, Morphogenesis physiology, N-Acetylglucosaminyltransferases genetics, Up-Regulation, Virulence genetics, Hepacivirus pathogenicity, Hepatitis C, Chronic genetics, N-Acetylglucosaminyltransferases physiology
- Abstract
Objective: Infection of human hepatocytes by the hepatitis C virus (HCV) is a multistep process involving both viral and host factors. microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression. Given that miRNAs were indicated to regulate between 30% and 75% of all human genes, we aimed to investigate the functional and regulatory role of miRNAs for the HCV life cycle., Design: To systematically reveal human miRNAs affecting the HCV life cycle, we performed a two-step functional high-throughput miRNA mimic screen in Huh7.5.1 cells infected with recombinant cell culture-derived HCV. miRNA targeting was then assessed using a combination of computational and functional approaches., Results: We uncovered miR-501-3p and miR-619-3p as novel modulators of HCV assembly/release. We discovered that these miRNAs regulate O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) protein expression and identified OGT and O-GlcNAcylation as regulators of HCV morphogenesis and infectivity. Furthermore, increased OGT expression in patient-derived liver tissue was associated with HCV-induced liver disease and cancer., Conclusion: miR-501-3p and miR-619-3p and their target OGT are previously undiscovered regulatory host factors for HCV assembly and infectivity. In addition to its effect on HCV morphogenesis, OGT may play a role in HCV-induced liver disease and hepatocarcinogenesis., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)
- Published
- 2020
- Full Text
- View/download PDF
12. Complete Genome Sequence and Annotation of the Staphylococcus aureus Strain HG001.
- Author
-
Caldelari I, Chane-Woon-Ming B, Noirot C, Moreau K, Romby P, Gaspin C, and Marzi S
- Abstract
Staphylococcus aureus is an opportunistic Gram-positive pathogen responsible for a wide range of infections from minor skin abscesses to life-threatening diseases. Here, we report the draft genome assembly and current annotation of the HG001 strain, a derivative of the RN1 (NCT8325) strain with restored rbsU (a positive activator of SigB)., (Copyright © 2017 Caldelari et al.)
- Published
- 2017
- Full Text
- View/download PDF
13. Identification of factors involved in target RNA-directed microRNA degradation.
- Author
-
Haas G, Cetin S, Messmer M, Chane-Woon-Ming B, Terenzi O, Chicher J, Kuhn L, Hammann P, and Pfeffer S
- Subjects
- Animals, Argonaute Proteins genetics, Argonaute Proteins metabolism, Base Sequence, Biotinylation, Cell Line, Tumor, Cytomegalovirus genetics, Cytomegalovirus Infections genetics, Cytomegalovirus Infections virology, Exoribonucleases metabolism, Gene Expression Regulation, HEK293 Cells, HeLa Cells, Hepatocytes cytology, Hepatocytes metabolism, Humans, Mice, MicroRNAs antagonists & inhibitors, MicroRNAs metabolism, Molecular Sequence Data, Nucleotidyltransferases metabolism, Oligonucleotides, Antisense genetics, Oligonucleotides, Antisense metabolism, Polynucleotide Adenylyltransferase, RNA, Messenger metabolism, mRNA Cleavage and Polyadenylation Factors genetics, mRNA Cleavage and Polyadenylation Factors metabolism, Exoribonucleases genetics, MicroRNAs genetics, Nucleotidyltransferases genetics, RNA Stability, RNA, Messenger genetics
- Abstract
The mechanism by which micro (mi)RNAs control their target gene expression is now well understood. It is however less clear how the level of miRNAs themselves is regulated. Under specific conditions, abundant and highly complementary target RNA can trigger miRNA degradation by a mechanism involving nucleotide addition and exonucleolytic degradation. One such mechanism has been previously observed to occur naturally during viral infection. To date, the molecular details of this phenomenon are not known. We report here that both the degree of complementarity and the ratio of miRNA/target abundance are crucial for the efficient decay of the small RNA. Using a proteomic approach based on the transfection of biotinylated antimiRNA oligonucleotides, we set to identify the factors involved in target-mediated miRNA degradation. Among the retrieved proteins, we identified members of the RNA-induced silencing complex, but also RNA modifying and degradation enzymes. We further validate and characterize the importance of one of these, the Perlman Syndrome 3'-5' exonuclease DIS3L2. We show that this protein interacts with Argonaute 2 and functionally validate its role in target-directed miRNA degradation both by artificial targets and in the context of mouse cytomegalovirus infection., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2016
- Full Text
- View/download PDF
14. Modulation of MicroRNA Cluster miR-183-96-182 Expression by Epstein-Barr Virus Latent Membrane Protein 1.
- Author
-
Oussaief L, Fendri A, Chane-Woon-Ming B, Poirey R, Delecluse HJ, Joab I, and Pfeffer S
- Subjects
- Base Sequence, Blotting, Northern, Cloning, Molecular, DNA Primers genetics, Gene Expression Profiling, Gene Knockout Techniques, Humans, MicroRNAs genetics, Molecular Sequence Data, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, RNA, Gene Expression Regulation genetics, MicroRNAs metabolism, Multigene Family genetics, Viral Matrix Proteins genetics
- Abstract
Unlabelled: Epstein-Barr virus (EBV) is an oncogenic human herpesvirus involved in the pathogenesis of Burkitt's lymphoma (BL) and various other lymphoproliferative disorders. In BL, EBV protein expression is restricted to EBV nuclear antigen 1 (EBNA1), but small noncoding RNAs such as EBV-encoded small RNAs (EBERs) and microRNAs (miRNAs) can also be detected. miRNAs play major roles in crucial processes such as proliferation, differentiation, and cell death. It has recently become clear that alterations in the expression profile of miRNAs contribute to the pathogenesis of a number of malignancies. During latent infection, EBV expresses 25 viral pre-miRNAs and modulates the expression of specific cellular miRNAs, such as miR-155 and miR-146, which potentially play a role in oncogenesis. Here, we established the small-RNA expression profiles of three BL cell lines. Using large-scale sequencing coupled to Northern blotting and real-time reverse transcription-PCR (RT-PCR) analysis validation, we demonstrated the differential expression of some cellular and viral miRNAs. High-level expression of the miR-183-96-182 cluster and EBV miR-BamHI A rightward transcript (miR-BART) cluster was significantly associated with EBV type I latency. This expression was not affected by viral reactivation since transforming growth factor β1 (TGF-β1) stimulation did not significantly change the miRNA profiles. However, using several approaches, including de novo infection with a mutant virus, we present evidence that the expression of latent membrane protein 1 (LMP-1) triggered downregulation of the expression of the miR-183-96-182 cluster. We further show that this effect involves the Akt signaling pathway., Importance: In addition to expressing their own miRNAs, herpesviruses also impact the expression levels of cellular miRNAs. This regulation can be either positive or negative and usually results in the perturbation of pathways to create a cellular environment that is more "virus-friendly." For example, EBV induces the expression of miR-155, a well-characterized oncomiR, which leads to increased cell proliferation and decreased cell death. Here, we show that EBV-encoded LMP-1 is also involved in the downregulation of a cluster of three miRNAs, miR-183, -96, and -182, which are known to be also repressed in several cancers. We therefore identify yet another potential player in EBV-induced oncogenesis., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
- Full Text
- View/download PDF
15. Cross-species comparative analysis of Dicer proteins during Sindbis virus infection.
- Author
-
Girardi E, Lefèvre M, Chane-Woon-Ming B, Paro S, Claydon B, Imler JL, Meignin C, and Pfeffer S
- Subjects
- Alphavirus Infections genetics, Animals, Animals, Genetically Modified, Cell Line, Drosophila, Gene Expression, Gene Silencing, High-Throughput Nucleotide Sequencing, Humans, Interferons metabolism, RNA Interference, RNA, Small Untranslated genetics, Ribonuclease III genetics, Signal Transduction, Alphavirus Infections metabolism, Alphavirus Infections virology, Ribonuclease III metabolism, Sindbis Virus genetics
- Abstract
In plants and invertebrates RNA silencing is a major defense mechanism against virus infections. The first event in RNA silencing is dicing of long double stranded RNAs into small interfering RNAs (siRNAs). The Dicer proteins involved in this process are phylogenetically conserved and have the same domain organization. Accordingly, the production of viral derived siRNAs has also been observed in the mouse, but only in restricted cell types. To gain insight on this restriction, we compare the dicing activity of human Dicer and fly Dicer-2 in the context of Sindbis virus (SINV) infection. Expression of human Dicer in flies inefficiently rescues the production of viral siRNAs but confers some protection against SINV. Conversely, expression of Dicer-2 in human cells allows the production of viral 21 nt small RNAs. However, this does not confer resistance to viral infection, but on the contrary results in stronger accumulation of viral RNA. We further show that Dicer-2 expression in human cells perturbs interferon (IFN) signaling pathways and antagonizes protein kinase R (PKR)-mediated antiviral immunity. Overall, our data suggest that a functional incompatibility between the Dicer and IFN pathways explains the predominance of the IFN response in mammalian somatic cells.
- Published
- 2015
- Full Text
- View/download PDF
16. Clearance of persistent hepatitis C virus infection in humanized mice using a claudin-1-targeting monoclonal antibody.
- Author
-
Mailly L, Xiao F, Lupberger J, Wilson GK, Aubert P, Duong FHT, Calabrese D, Leboeuf C, Fofana I, Thumann C, Bandiera S, Lütgehetmann M, Volz T, Davis C, Harris HJ, Mee CJ, Girardi E, Chane-Woon-Ming B, Ericsson M, Fletcher N, Bartenschlager R, Pessaux P, Vercauteren K, Meuleman P, Villa P, Kaderali L, Pfeffer S, Heim MH, Neunlist M, Zeisel MB, Dandri M, McKeating JA, Robinet E, and Baumert TF
- Subjects
- Animals, Antibodies, Monoclonal immunology, Antibodies, Monoclonal, Humanized administration & dosage, Antibodies, Monoclonal, Humanized immunology, Claudin-1 therapeutic use, Hepacivirus immunology, Hepacivirus pathogenicity, Hepatitis C immunology, Hepatitis C virology, Hepatocytes immunology, Humans, Liver Cirrhosis therapy, Liver Cirrhosis virology, Mice, Antibodies, Monoclonal administration & dosage, Claudin-1 immunology, Hepatitis C therapy, Liver Cirrhosis immunology
- Abstract
Hepatitis C virus (HCV) infection is a leading cause of liver cirrhosis and cancer. Cell entry of HCV and other pathogens is mediated by tight junction (TJ) proteins, but successful therapeutic targeting of TJ proteins has not been reported yet. Using a human liver-chimeric mouse model, we show that a monoclonal antibody specific for the TJ protein claudin-1 (ref. 7) eliminates chronic HCV infection without detectable toxicity. This antibody inhibits HCV entry, cell-cell transmission and virus-induced signaling events. Antibody treatment reduces the number of HCV-infected hepatocytes in vivo, highlighting the need for de novo infection by means of host entry factors to maintain chronic infection. In summary, we demonstrate that an antibody targeting a virus receptor can cure chronic viral infection and uncover TJ proteins as targets for antiviral therapy.
- Published
- 2015
- Full Text
- View/download PDF
17. Small RNA cloning and sequencing strategy affects host and viral microRNA expression signatures.
- Author
-
Stik G, Muylkens B, Coupeau D, Laurent S, Dambrine G, Messmer M, Chane-Woon-Ming B, Pfeffer S, and Rasschaert D
- Subjects
- Animals, Chickens genetics, Chickens virology, High-Throughput Nucleotide Sequencing, Marek Disease genetics, Marek Disease virology, MicroRNAs genetics, Transcriptome, Herpesvirus 2, Gallid genetics, MicroRNAs biosynthesis, RNA, Viral biosynthesis
- Abstract
The establishment of the microRNA (miRNA) expression signatures is the basic element to investigate the role played by these regulatory molecules in the biology of an organism. Marek's disease virus 1 (MDV-1) is an avian herpesvirus that naturally infects chicken and induces T cells lymphomas. During latency, MDV-1, like other herpesviruses, expresses a limited subset of transcripts. These include three miRNA clusters. Several studies identified the expression of virus and host encoded miRNAs from MDV-1 infected cell cultures and chickens. But a high discrepancy was observed when miRNA cloning frequencies obtained from different cloning and sequencing protocols were compared. Thus, we analyzed the effect of small RNA library preparation and sequencing on the miRNA frequencies obtained from the same RNA samples collected during MDV-1 infection of chicken at different steps of the oncoviral pathogenesis. Qualitative and quantitative variations were found in the data, depending on the strategy used. One of the mature miRNA derived from the latency-associated-transcript (LAT), mdv1-miR-M7-5p, showed the highest variation. Its cloning frequency was 50% of the viral miRNA counts when a small scale sequencing approach was used. Its frequency was 100 times less abundant when determined through the deep sequencing approach. Northern blot analysis showed a better correlation with the miRNA frequencies found by the small scale sequencing approach. By analyzing the cellular miRNA repertoire, we also found a gap between the two sequencing approaches. Collectively, our study indicates that next-generation sequencing data considered alone are limited for assessing the absolute copy number of transcripts. Thus, the quantification of small RNA should be addressed by compiling data obtained by using different techniques such as microarrays, qRT-PCR and NB analysis in support of high throughput sequencing data. These observations should be considered when miRNA variations are studied prior addressing functional studies., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
18. Importance of the RNA secondary structure for the relative accumulation of clustered viral microRNAs.
- Author
-
Contrant M, Fender A, Chane-Woon-Ming B, Randrianjafy R, Vivet-Boudou V, Richer D, and Pfeffer S
- Subjects
- B-Lymphocytes virology, Cell Line, Herpesvirus 8, Human genetics, Humans, MicroRNAs metabolism, Mutagenesis, Nucleic Acid Conformation, RNA, Viral metabolism, MicroRNAs chemistry, RNA, Viral chemistry
- Abstract
Micro (mi)RNAs are small non-coding RNAs with key regulatory functions. Recent advances in the field allowed researchers to identify their targets. However, much less is known regarding the regulation of miRNAs themselves. The accumulation of these tiny regulators can be modulated at various levels during their biogenesis from the transcription of the primary transcript (pri-miRNA) to the stability of the mature miRNA. Here, we studied the importance of the pri-miRNA secondary structure for the regulation of mature miRNA accumulation. To this end, we used the Kaposi's sarcoma herpesvirus, which encodes a cluster of 12 pre-miRNAs. Using small RNA profiling and quantitative northern blot analysis, we measured the absolute amount of each mature miRNAs in different cellular context. We found that the difference in expression between the least and most expressed viral miRNAs could be as high as 60-fold. Using high-throughput selective 2'-hydroxyl acylation analyzed by primer extension, we then determined the secondary structure of the long primary transcript. We found that highly expressed miRNAs derived from optimally structured regions within the pri-miRNA. Finally, we confirmed the importance of the local structure by swapping stem-loops or by targeted mutagenesis of selected miRNAs, which resulted in a perturbed accumulation of the mature miRNA., (© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2014
- Full Text
- View/download PDF
19. Identification of RNase L-dependent, 3'-end-modified, viral small RNAs in Sindbis virus-infected mammalian cells.
- Author
-
Girardi E, Chane-Woon-Ming B, Messmer M, Kaukinen P, and Pfeffer S
- Subjects
- Animals, Cell Line, Chlorocebus aethiops, Gene Expression Profiling, Humans, RNA Processing, Post-Transcriptional, Endoribonucleases metabolism, Host-Pathogen Interactions, RNA, Small Untranslated metabolism, RNA, Viral metabolism, Sindbis Virus physiology
- Abstract
Unlabelled: Small RNAs play a critical role in host-pathogen interaction. Indeed, small RNA-mediated silencing or RNA interference (RNAi) is one of the earliest forms of antiviral immunity. Although it represents the main defense system against viruses in many organisms, the antiviral role of RNAi has not been clearly proven in higher vertebrates. However, it is well established that their response to viral infection relies on the recognition of viral RNAs by host pattern recognition receptors (PRRs) to trigger activation of the interferon pathway. In the present work, we report the existence of a novel small noncoding RNA population produced in mammalian cells upon RNA virus infection. Using Sindbis virus (SINV) as a prototypic arbovirus model, we profiled the small RNA population of infected cells in both human and African green monkey cell lines. Here, we provide evidence for the presence of discrete small RNAs of viral origin that are not associated with the RNA-induced silencing complex (RISC), that are highly expressed and detected by Northern blot analysis, and that accumulate as 21- to 28-nucleotide (nt) species during infection. We report that the cellular antiviral endoribonuclease RNase L cleaves the viral genome, producing in turn the small RNAs. Surprisingly, we uncovered the presence of a modification on the 3'-end nucleotide of SINV-derived viral small RNAs (SvsRNAs) that might be at the origin of their stability. Altogether, our findings show that stable modified small viral RNAs could represent a novel way to modulate host-virus interaction upon SINV infection., Importance: In a continuous arms race, viruses have to deal with host antiviral responses in order to successfully establish an infection. In mammalian cells, the host defense mechanism relies on the recognition of viral RNAs, resulting in the activation of type I interferons (IFNs). In turn, the expression of many interferon-stimulated genes (ISGs) is induced to inhibit viral replication. Here we report that the cytoplasmic, interferon-induced, cellular endoribonuclease RNase L is involved in the accumulation of a novel small RNA population of viral origin. These small RNAs are produced upon SINV infection of mammalian cells and are stabilized by a 3'-end modification. Altogether, our findings indicate that in our system RNA silencing is not active against Sindbis virus (SINV) and might open the way to a better understanding of the antiviral response mediated by a novel class of small RNAs.
- Published
- 2013
- Full Text
- View/download PDF
20. Mutation rate dynamics in a bacterial population reflect tension between adaptation and genetic load.
- Author
-
Wielgoss S, Barrick JE, Tenaillon O, Wiser MJ, Dittmar WJ, Cruveiller S, Chane-Woon-Ming B, Médigue C, Lenski RE, and Schneider D
- Subjects
- DNA Glycosylases genetics, Escherichia coli Proteins genetics, Likelihood Functions, Models, Genetic, Phylogeny, Population Dynamics, Pyrophosphatases genetics, Adaptation, Biological genetics, Biological Evolution, Escherichia coli genetics, Genetic Load, Mutation Rate
- Abstract
Mutations are the ultimate source of heritable variation for evolution. Understanding how mutation rates themselves evolve is thus essential for quantitatively understanding many evolutionary processes. According to theory, mutation rates should be minimized for well-adapted populations living in stable environments, whereas hypermutators may evolve if conditions change. However, the long-term fate of hypermutators is unknown. Using a phylogenomic approach, we found that an adapting Escherichia coli population that first evolved a mutT hypermutator phenotype was later invaded by two independent lineages with mutY mutations that reduced genome-wide mutation rates. Applying neutral theory to synonymous substitutions, we dated the emergence of these mutations and inferred that the mutT mutation increased the point-mutation rate by ∼150-fold, whereas the mutY mutations reduced the rate by ∼40-60%, with a corresponding decrease in the genetic load. Thus, the long-term fate of the hypermutators was governed by the selective advantage arising from a reduced mutation rate as the potential for further adaptation declined.
- Published
- 2013
- Full Text
- View/download PDF
21. Degradation of cellular mir-27 by a novel, highly abundant viral transcript is important for efficient virus replication in vivo.
- Author
-
Marcinowski L, Tanguy M, Krmpotic A, Rädle B, Lisnić VJ, Tuddenham L, Chane-Woon-Ming B, Ruzsics Z, Erhard F, Benkartek C, Babic M, Zimmer R, Trgovcich J, Koszinowski UH, Jonjic S, Pfeffer S, and Dölken L
- Subjects
- Animals, Argonaute Proteins genetics, Argonaute Proteins metabolism, Binding Sites, Cell Line, Down-Regulation genetics, Gene Expression Regulation genetics, High-Throughput Nucleotide Sequencing, Humans, Mice, Mice, Inbred BALB C, Mice, SCID, MicroRNAs genetics, Muromegalovirus genetics, Mutation, RNA Processing, Post-Transcriptional, RNA Stability genetics, RNA, Viral genetics, Sequence Analysis, RNA, 3' Untranslated Regions genetics, Cytomegalovirus Infections virology, MicroRNAs metabolism, Muromegalovirus physiology, RNA, Viral metabolism, Virus Replication genetics
- Abstract
Cytomegaloviruses express large amounts of viral miRNAs during lytic infection, yet, they only modestly alter the cellular miRNA profile. The most prominent alteration upon lytic murine cytomegalovirus (MCMV) infection is the rapid degradation of the cellular miR-27a and miR-27b. Here, we report that this regulation is mediated by the ∼1.7 kb spliced and highly abundant MCMV m169 transcript. Specificity to miR-27a/b is mediated by a single, apparently optimized, miRNA binding site located in its 3'-UTR. This site is easily and efficiently retargeted to other cellular and viral miRNAs by target site replacement. Expression of the 3'-UTR of m169 by an adenoviral vector was sufficient to mediate its function, indicating that no other viral factors are essential in this process. Degradation of miR-27a/b was found to be accompanied by 3'-tailing and -trimming. Despite its dramatic effect on miRNA stability, we found this interaction to be mutual, indicating potential regulation of m169 by miR-27a/b. Most interestingly, three mutant viruses no longer able to target miR-27a/b, either due to miRNA target site disruption or target site replacement, showed significant attenuation in multiple organs as early as 4 days post infection, indicating that degradation of miR-27a/b is important for efficient MCMV replication in vivo.
- Published
- 2012
- Full Text
- View/download PDF
22. Small RNA deep sequencing identifies microRNAs and other small noncoding RNAs from human herpesvirus 6B.
- Author
-
Tuddenham L, Jung JS, Chane-Woon-Ming B, Dölken L, and Pfeffer S
- Subjects
- Base Sequence, Cell Line, DNA Primers, Fluorescent Antibody Technique, Genes, Viral, Humans, Likelihood Functions, Replication Origin, Reverse Transcriptase Polymerase Chain Reaction, Herpesvirus 6, Human genetics, MicroRNAs genetics, RNA, Untranslated genetics
- Abstract
Roseolovirus, or human herpesvirus 6 (HHV-6), is a ubiquitous human pathogen infecting over 95% of the population by the age of 2 years. As with other herpesviruses, reactivation of HHV-6 can present with severe complications in immunocompromised individuals. Recent studies have highlighted the importance of herpesvirus-derived microRNAs (miRNAs) in modulating both cellular and viral gene expression. An initial report which computed the likelihood of various viruses to encode miRNAs did not predict HHV-6 miRNAs. To experimentally screen for small HHV-6-encoded RNAs, we conducted large-scale sequencing of Sup-T-1 cells lytically infected with a laboratory strain of HHV-6B. This revealed an abundant, 60- to 65-nucleotide RNA of unknown function derived from the lytic origin of replication (OriLyt) that gave rise to smaller RNA species of 18 or 19 nucleotides. In addition, we identified four pre-miRNAs whose mature forms accumulated in Argonaute 2. In contrast to the case for other betaherpesviruses, HHV-6B miRNAs are expressed from direct repeat regions (DR(L) and DR(R)) located at either side of the genome. All miRNAs are conserved in the closely related HHV-6A variant, and one of them is a seed ortholog of the human miRNA miR-582-5p. Similar to alphaherpesvirus miRNAs, they are expressed in antisense orientation relative to immediate-early open reading frames (ORFs) and thus have the potential to regulate key viral genes.
- Published
- 2012
- Full Text
- View/download PDF
23. Metabolic diversity among main microorganisms inside an arsenic-rich ecosystem revealed by meta- and proteo-genomics.
- Author
-
Bertin PN, Heinrich-Salmeron A, Pelletier E, Goulhen-Chollet F, Arsène-Ploetze F, Gallien S, Lauga B, Casiot C, Calteau A, Vallenet D, Bonnefoy V, Bruneel O, Chane-Woon-Ming B, Cleiss-Arnold J, Duran R, Elbaz-Poulichet F, Fonknechten N, Giloteaux L, Halter D, Koechler S, Marchal M, Mornico D, Schaeffer C, Smith AA, Van Dorsselaer A, Weissenbach J, Médigue C, and Le Paslier D
- Subjects
- Bacteria classification, Bacteria isolation & purification, Iron metabolism, Mining, Phylogeny, Reverse Transcriptase Polymerase Chain Reaction, Sulfur metabolism, Arsenic metabolism, Bacteria genetics, Bacteria metabolism, Ecosystem, Metagenomics, Proteomics
- Abstract
By their metabolic activities, microorganisms have a crucial role in the biogeochemical cycles of elements. The complete understanding of these processes requires, however, the deciphering of both the structure and the function, including synecologic interactions, of microbial communities. Using a metagenomic approach, we demonstrated here that an acid mine drainage highly contaminated with arsenic is dominated by seven bacterial strains whose genomes were reconstructed. Five of them represent yet uncultivated bacteria and include two strains belonging to a novel bacterial phylum present in some similar ecosystems, and which was named 'Candidatus Fodinabacter communificans.' Metaproteomic data unravelled several microbial capabilities expressed in situ, such as iron, sulfur and arsenic oxidation that are key mechanisms in biomineralization, or organic nutrient, amino acid and vitamin metabolism involved in synthrophic associations. A statistical analysis of genomic and proteomic data and reverse transcriptase-PCR experiments allowed us to build an integrated model of the metabolic interactions that may be of prime importance in the natural attenuation of such anthropized ecosystems.
- Published
- 2011
- Full Text
- View/download PDF
24. Mutation Rate Inferred From Synonymous Substitutions in a Long-Term Evolution Experiment With Escherichia coli.
- Author
-
Wielgoss S, Barrick JE, Tenaillon O, Cruveiller S, Chane-Woon-Ming B, Médigue C, Lenski RE, and Schneider D
- Abstract
The quantification of spontaneous mutation rates is crucial for a mechanistic understanding of the evolutionary process. In bacteria, traditional estimates using experimental or comparative genetic methods are prone to statistical uncertainty and consequently estimates vary by over one order of magnitude. With the advent of next-generation sequencing, more accurate estimates are now possible. We sequenced 19 Escherichia coli genomes from a 40,000-generation evolution experiment and directly inferred the point-mutation rate based on the accumulation of synonymous substitutions. The resulting estimate was 8.9 × 10(-11) per base-pair per generation, and there was a significant bias toward increased AT-content. We also compared our results with published genome sequence datasets for other bacterial evolution experiments. Given the power of our approach, our estimate represents the most accurate measure of bacterial base-substitution rates available to date.
- Published
- 2011
- Full Text
- View/download PDF
25. Molecular and evolutionary bases of within-patient genotypic and phenotypic diversity in Escherichia coli extraintestinal infections.
- Author
-
Levert M, Zamfir O, Clermont O, Bouvet O, Lespinats S, Hipeaux MC, Branger C, Picard B, Saint-Ruf C, Norel F, Balliau T, Zivy M, Le Nagard H, Cruveiller S, Chane-Woon-Ming B, Nilsson S, Gudelj I, Phan K, Ferenci T, Tenaillon O, and Denamur E
- Subjects
- Adult, Animals, Bacterial Proteins genetics, Bacterial Proteins metabolism, Cell Movement, DNA, Bacterial genetics, Electrophoresis, Gel, Two-Dimensional, Escherichia coli classification, Escherichia coli genetics, Escherichia coli Infections epidemiology, Escherichia coli Infections genetics, Female, Genome, Bacterial, Genotype, Humans, Hydrogen Peroxide pharmacology, Immunoblotting, Mice, Microbial Sensitivity Tests, Models, Theoretical, Mutation genetics, Oxidants pharmacology, Polymerase Chain Reaction, Sigma Factor genetics, Sigma Factor metabolism, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Virulence Factors genetics, Anti-Bacterial Agents pharmacology, Biological Evolution, Drug Resistance, Bacterial genetics, Escherichia coli pathogenicity, Escherichia coli Infections microbiology, Escherichia coli Proteins genetics, Genetic Variation, Virulence genetics
- Abstract
Although polymicrobial infections, caused by combinations of viruses, bacteria, fungi and parasites, are being recognised with increasing frequency, little is known about the occurrence of within-species diversity in bacterial infections and the molecular and evolutionary bases of this diversity. We used multiple approaches to study the genomic and phenotypic diversity among 226 Escherichia coli isolates from deep and closed visceral infections occurring in 19 patients. We observed genomic variability among isolates from the same site within 11 patients. This diversity was of two types, as patients were infected either by several distinct E. coli clones (4 patients) or by members of a single clone that exhibit micro-heterogeneity (11 patients); both types of diversity were present in 4 patients. A surprisingly wide continuum of antibiotic resistance, outer membrane permeability, growth rate, stress resistance, red dry and rough morphotype characteristics and virulence properties were present within the isolates of single clones in 8 of the 11 patients showing genomic micro-heterogeneity. Many of the observed phenotypic differences within clones affected the trade-off between self-preservation and nutritional competence (SPANC). We showed in 3 patients that this phenotypic variability was associated with distinct levels of RpoS in co-existing isolates. Genome mutational analysis and global proteomic comparisons in isolates from a patient revealed a star-like relationship of changes amongst clonally diverging isolates. A mathematical model demonstrated that multiple genotypes with distinct RpoS levels can co-exist as a result of the SPANC trade-off. In the cases involving infection by a single clone, we present several lines of evidence to suggest diversification during the infectious process rather than an infection by multiple isolates exhibiting a micro-heterogeneity. Our results suggest that bacteria are subject to trade-offs during an infectious process and that the observed diversity resembled results obtained in experimental evolution studies. Whatever the mechanisms leading to diversity, our results have strong medical implications in terms of the need for more extensive isolate testing before deciding on antibiotic therapies.
- Published
- 2010
- Full Text
- View/download PDF
26. Delineation and analysis of chromosomal regions specifying Yersinia pestis.
- Author
-
Derbise A, Chenal-Francisque V, Huon C, Fayolle C, Demeure CE, Chane-Woon-Ming B, Médigue C, Hinnebusch BJ, and Carniel E
- Subjects
- Animals, Female, Mice, Open Reading Frames, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Virulence, Yersinia pestis pathogenicity, Chromosome Mapping, Yersinia pestis genetics
- Abstract
Yersinia pestis, the causative agent of plague, has recently diverged from the less virulent enteropathogen Yersinia pseudotuberculosis. Its emergence has been characterized by massive genetic loss and inactivation and limited gene acquisition. The acquired genes include two plasmids, a filamentous phage, and a few chromosomal loci. The aim of this study was to characterize the chromosomal regions acquired by Y. pestis. Following in silico comparative analysis and PCR screening of 98 strains of Y. pseudotuberculosis and Y. pestis, we found that eight chromosomal loci (six regions [R1pe to R6pe] and two coding sequences [CDS1pe and CDS2pe]) specified Y. pestis. Signatures of integration by site specific or homologous recombination were identified for most of them. These acquisitions and the loss of ancestral DNA sequences were concentrated in a chromosomal region opposite to the origin of replication. The specific regions were acquired very early during Y. pestis evolution and were retained during its microevolution, suggesting that they might bring some selective advantages. Only one region (R3pe), predicted to carry a lambdoid prophage, is most likely no longer functional because of mutations. With the exception of R1pe and R2pe, which have the potential to encode a restriction/modification and a sugar transport system, respectively, no functions could be predicted for the other Y. pestis-specific loci. To determine the role of the eight chromosomal loci in the physiology and pathogenicity of the plague bacillus, each of them was individually deleted from the bacterial chromosome. None of the deletants exhibited defects during growth in vitro. Using the Xenopsylla cheopis flea model, all deletants retained the capacity to produce a stable and persistent infection and to block fleas. Similarly, none of the deletants caused any acute flea toxicity. In the mouse model of infection, all deletants were fully virulent upon subcutaneous or aerosol infections. Therefore, our results suggest that acquisition of new chromosomal materials has not been of major importance in the dramatic change of life cycle that has accompanied the emergence of Y. pestis.
- Published
- 2010
- Full Text
- View/download PDF
27. Experimental evolution of a plant pathogen into a legume symbiont.
- Author
-
Marchetti M, Capela D, Glew M, Cruveiller S, Chane-Woon-Ming B, Gris C, Timmers T, Poinsot V, Gilbert LB, Heeb P, Médigue C, Batut J, and Masson-Boivin C
- Subjects
- Adaptation, Biological, Chimera, Directed Molecular Evolution, Gene Transfer, Horizontal, Nitrogen Fixation, Plant Root Nodulation genetics, Polymorphism, Single Nucleotide, Rhizobium physiology, Fabaceae microbiology, Rhizobium genetics, Symbiosis genetics
- Abstract
Rhizobia are phylogenetically disparate alpha- and beta-proteobacteria that have achieved the environmentally essential function of fixing atmospheric nitrogen in symbiosis with legumes. Ample evidence indicates that horizontal transfer of symbiotic plasmids/islands has played a crucial role in rhizobia evolution. However, adaptive mechanisms that allow the recipient genomes to express symbiotic traits are unknown. Here, we report on the experimental evolution of a pathogenic Ralstonia solanacearum chimera carrying the symbiotic plasmid of the rhizobium Cupriavidus taiwanensis into Mimosa nodulating and infecting symbionts. Two types of adaptive mutations in the hrpG-controlled virulence pathway of R. solanacearum were identified that are crucial for the transition from pathogenicity towards mutualism. Inactivation of the hrcV structural gene of the type III secretion system allowed nodulation and early infection to take place, whereas inactivation of the master virulence regulator hrpG allowed intracellular infection of nodule cells. Our findings predict that natural selection of adaptive changes in the legume environment following horizontal transfer has been a major driving force in rhizobia evolution and diversification and show the potential of experimental evolution to decipher the mechanisms leading to symbiosis., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2010
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.