Comparative Exoproteome Analysis of Streptococcus suis Human Isolates

The swine pathogen Streptococcus suis is a Gram-positive bacterium which causes infections in pigs, with an impact in animal health and in the livestock industry, and it is also an important zoonotic agent. During the infection process, surface and secreted proteins are essential in the interaction between microorganisms and their hosts. Here, we report a comparative proteomic analysis of the proteins released to the extracellular milieu in six human clinical isolates belonging to the highly prevalent and virulent serotype 2. The total secreted content was precipitated and analyzed by GeLC-MS/MS. In the six strains, 144 proteins assigned to each of the categories of extracellular or surface proteins were identified, as well as 680 predicted cytoplasmic proteins, many of which are putative moonlighting proteins. Of the nine predicted signal peptide-I secreted proteins, seven had relevant antigenic potential when they were analyzed through bioinformatic analysis. This is the first work comparing the exoproteome fraction of several human isolates of this important pathogen.

Download Full-text

Proteomic and Bioinformatic Analysis of Streptococcus suis Human Isolates: Combined Prediction of Potential Vaccine Candidates

Vaccines ◽

10.3390/vaccines8020188 ◽

2020 ◽

Vol 8 (2) ◽

pp. 188 ◽

Cited By ~ 2

Author(s):

Esther Prados de la Torre ◽

Antonio Rodríguez-Franco ◽

Manuel J. Rodríguez-Ortega

Keyword(s):

Streptococcus Suis ◽

Bioinformatic Analysis ◽

Surface Proteins ◽

Clinical Isolates ◽

Economic Losses ◽

Bacterial Cells ◽

Label Free ◽

Bioinformatic Tools ◽

Potential Vaccine ◽

Live Bacterial Cells

Streptococcus suis is a Gram-positive bacterium responsible for major infections in pigs and economic losses in the livestock industry, but also an emerging zoonotic pathogen causing serious diseases in humans. No vaccine is available so far against this microorganism. Conserved surface proteins are among the most promising candidates for new and effective vaccines. Until now, research on this pathogen has focused on swine isolates, but there is a lack of studies to identify and characterize surface proteins from human clinical isolates. In this work, we performed a comparative proteomic analysis of six clinical isolates from human patients, all belonging to the major serotype 2, by “shaving” the live bacterial cells with trypsin, followed by LC-MS/MS analysis. We identified 131 predicted surface proteins and carried out a label-free semi-quantitative analysis of protein abundances within the six strains. Then, we combined our proteomics results with bioinformatic tools to help improving the selection of novel antigens that can enter the pipeline of vaccine candidate testing. Our work is then a complement to the reverse vaccinology concept.

Download Full-text

Translation of hepatitis B virus (HBV) surface proteins from the HBV pregenome and precore RNAs in Semliki Forest virus-driven expression

Journal of General Virology ◽

10.1099/vir.0.80388-0 ◽

2004 ◽

Vol 85 (11) ◽

pp. 3343-3351 ◽

Cited By ~ 6

Author(s):

Anna Zajakina ◽

Tatyana Kozlovska ◽

Ruta Bruvere ◽

Jekaterina Aleksejeva ◽

Paul Pumpens ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Semliki Forest Virus ◽

Expression System ◽

Protein A ◽

Direct Repeat ◽

Infection Process ◽

Surface Proteins ◽

B Virus ◽

Internal Translation

Hepatitis B virus (HBV) pregenome RNA (pgRNA) serves as a translation template for the HBV core (HBc) protein and viral polymerase (Pol). HBV precore RNA (pcRNA) directs the synthesis of the precore (preC) protein, a precursor of the hepatitis B e antigen (HBeAg). pgRNA and pcRNA were expressed in the Semliki Forest virus (SFV) expression system. Besides the HBc and preC proteins, there was revealed the synthesis of all three forms of HBV surface (HBs) proteins: long (LHBs), middle (MHBs) and short (SHBs), the start codons of which are located more than 1000 nt downstream of the HBc and preC start codons. Moreover, other HBV templates, such as 3′-truncated pgRNA lacking 3′ direct repeat and Pol mRNA, both carrying internally the HBs sequences, provided the synthesis of three HBs protein forms in the SFV-driven expression system. Maximal production of the HBs was provided by Pol mRNA, while HBc- and preC-producing templates showed relatively low internal translation of the HBs. These data allow the proposal of a ribosome leaky scanning model of internal translation initiation for HBs proteins. The putative functional role of such exceptional synthesis of the HBs proteins from the pgRNA and pcRNA templates in the natural HBV infection process needs further evaluation.

Download Full-text

ETosis: A Microbicidal Mechanism beyond Cell Death

Journal of Parasitology Research ◽

10.1155/2012/929743 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 79

Author(s):

Anderson B. Guimarães-Costa ◽

Michelle T. C. Nascimento ◽

Amanda B. Wardini ◽

Lucia H. Pinto-da-Silva ◽

Elvira M. Saraiva

Keyword(s):

Cell Death ◽

Mast Cells ◽

Cell Types ◽

Neutrophil Extracellular Traps ◽

Protozoan Parasites ◽

Extracellular Milieu ◽

Extracellular Traps ◽

Cytoplasmic Proteins

Netosis is a recently described type of neutrophil death occurring with the release to the extracellular milieu of a lattice composed of DNA associated with histones and granular and cytoplasmic proteins. These webs, initially named neutrophil extracellular traps (NETs), ensnare and kill microorganisms. Similarly, other cell types, such as eosinophils, mast cells, and macrophages, can also dye by this mechanism; thus, it was renamed as ETosis, meaning death with release of extracellular traps (ETs). Here, we review the mechanism of NETosis/etosis, emphasizing its role in diseases caused by protozoan parasites, fungi, and viruses.

Download Full-text

Characterization and Protective Activity of Monoclonal Antibodies Directed against Streptococcus suis Serotype 2 Capsular Polysaccharide Obtained Using a Glycoconjugate

Pathogens ◽

10.3390/pathogens8030139 ◽

2019 ◽

Vol 8 (3) ◽

pp. 139 ◽

Cited By ~ 1

Author(s):

Guillaume Goyette-Desjardins ◽

Sonia Lacouture ◽

Jean-Philippe Auger ◽

René Roy ◽

Marcelo Gottschalk ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Capsular Polysaccharide ◽

Passive Immunization ◽

Streptococcus Suis ◽

Diagnostic Tools ◽

Protective Activity ◽

Lethal Challenge ◽

Swine Pathogen ◽

Suis Serotype ◽

Streptococcus Suis Serotype 2

Streptococcus suis serotype 2 is an encapsulated bacterium and an important swine pathogen. Opsonizing antibody responses targeting capsular polysaccharides (CPSs) are protective against extracellular pathogens. To elucidate the protective activity of monoclonal antibodies (mAbs) directed against S. suis serotype 2 CPS, mice were immunized with a serotype 2 CPS-glycoconjugate and three hybridomas were isolated; of which, two were murine IgMs and the other a murine IgG1. Whereas the IgMs (mAbs 9E7 and 13C8) showed different reactivity levels with S. suis serotypes 1, 1/2, 2 and 14, the IgG1 (mAb 16H11) was shown to be serotype 2-specific. All mAbs targeted the sialylated chain of the CPSs. Using an opsonophagocytosis assay, the IgMs were opsonizing towards the S. suis serotypes to which they cross-react, while the IgG1 failed to induce bacterial elimination. In a model of mouse passive immunization followed by a lethal challenge with S. suis serotype 2, the IgG1 and IgM cross-reacting only with serotype 14 (mAb 13C8) failed to protect, while the IgM cross-reacting with serotypes 1, 1/2, and 14 (mAb 9E7) was shown to be protective by limiting bacteremia. These new mAbs show promise as new S. suis diagnostic tools, as well as potential for therapeutic applications.

Download Full-text

Characteristic Dissection of Xanthomonas oryzae pv. oryzae Responsive MicroRNAs in Rice

International Journal of Molecular Sciences ◽

10.3390/ijms21030785 ◽

2020 ◽

Vol 21 (3) ◽

pp. 785

Author(s):

Yanfeng Jia ◽

Chunrong Li ◽

Quanlin Li ◽

Pengcheng Liu ◽

Dongfeng Liu ◽

...

Keyword(s):

Rice Variety ◽

Bioinformatic Analysis ◽

Infection Process ◽

Xanthomonas Oryzae ◽

Differentially Expressed ◽

Small Rna Sequencing ◽

Sequencing Data ◽

Stem Loop ◽

Differentially Expressed Mirnas ◽

Plant Pathogen Interaction

MicroRNAs (miRNAs) are crucial player in plant-pathogen interaction. While the evidence has demonstrated that rice miRNAs mediate immune response to pathogens invasion, the roles of miRNAs on Xanthomonas oryzae pv. oryzae (Xoo) attack remain be in place. Herein, we monitored the responsive changes of rice miRNAs at 0, 8, 24 h across Xoo strain PXO86 infection in its compatible rice variety IR24 and incompatible variety IRBB5 by small RNA sequencing, and the genes targeted by miRNAs were also detected via degradome technology. The faithfulness of sequencing data was validated through quantitative real-time stem-loop reverse transcription-polymerase chain reaction assay. Bioinformatic analysis showed that the differentially expressed miRNAs could be divided into three immunity-related clusters, and 80 regulatory units were emerged in infection process, which comprises 29 differentially expressed known miRNAs and 38 cleaved targets. Furthermore, the miRNA presumptive function of separate immunity cluster in rice-Xoo interplay was confirmed through overexpressing osa-miR164a, osa-miR167d and osa-miR159b, and the disruption of regulatory units, osa-miR164a/OsNAC60, osa-miR167d-5p/OsWD40-174 and osa-miR159b/OsMYBGA, OsLRR-RLK2, OsMPK20-4, may reset rice defense response to Xoo infestation in a controllable manner. These findings provide new insights into the complex roles of characteristic miRNAs and their targets in rice-Xoo interactions.

Download Full-text

Ubiquitous Carbohydrate Binding Modules Decorate 936 Lactococcal Siphophage Virions

Viruses ◽

10.3390/v11070631 ◽

2019 ◽

Vol 11 (7) ◽

pp. 631 ◽

Cited By ~ 7

Author(s):

Stephen Hayes ◽

Jennifer Mahony ◽

Renaud Vincentelli ◽

Laurie Ramond ◽

Arjen Nauta ◽

...

Keyword(s):

Fluorescent Protein ◽

Bioinformatic Analysis ◽

Infection Process ◽

Ecological Niches ◽

Carbohydrate Binding ◽

Binding Assays ◽

Protein Fusion ◽

Carbohydrate Binding Modules ◽

Structural Assembly ◽

Green Fluorescent

With the availability of an increasing number of 3D structures of bacteriophage components, combined with powerful in silico predictive tools, it has become possible to decipher the structural assembly and functionality of phage adhesion devices. In the current study, we examined 113 members of the 936 group of lactococcal siphophages, and identified a number of Carbohydrate Binding Modules (CBMs) in the neck passage structure and major tail protein, on top of evolved Dit proteins, as recently reported by us. The binding ability of such CBM-containing proteins was assessed through the construction of green fluorescent protein fusion proteins and subsequent binding assays. Two CBMs, one from the phage tail and another from the neck, demonstrated definite binding to their phage-specific host. Bioinformatic analysis of the structural proteins of 936 phages reveals that they incorporate binding modules which exhibit structural homology to those found in other lactococcal phage groups and beyond, indicating that phages utilize common structural “bricks” to enhance host binding capabilities. The omnipresence of CBMs in Siphophages supports their beneficial role in the infection process, as they can be combined in various ways to form appendages with different shapes and functionalities, ensuring their success in host detection in their respective ecological niches.

Download Full-text

Involvement of Various Enzymes in the Physiology and Pathogenesis of Streptococcus suis

Veterinary Sciences ◽

10.3390/vetsci7040143 ◽

2020 ◽

Vol 7 (4) ◽

pp. 143

Author(s):

Chengkun Zheng ◽

Man Wei ◽

Mengdie Jia ◽

ManMan Cao

Keyword(s):

Streptococcus Suis ◽

Infection Process ◽

Swine Industry ◽

Serine Threonine Kinase ◽

Future Directions ◽

Threonine Kinase ◽

Severe Infections ◽

Living Organisms ◽

Almost All

Streptococcus suis causes severe infections in both swine and humans, making it a serious threat to the swine industry and public health. Insight into the physiology and pathogenesis of S. suis undoubtedly contributes to the control of its infection. During the infection process, a wide variety of virulence factors enable S. suis to colonize, invade, and spread in the host, thus causing localized infections and/or systemic diseases. Enzymes catalyze almost all aspects of metabolism in living organisms. Numerous enzymes have been characterized in extensive detail in S. suis, and have shown to be involved in the pathogenesis and/or physiology of this pathogen. In this review, we describe the progress in the study of some representative enzymes in S. suis, such as ATPases, immunoglobulin-degrading enzymes, and eukaryote-like serine/threonine kinase and phosphatase, and we highlight the important role of various enzymes in the physiology and pathogenesis of this pathogen. The controversies about the current understanding of certain enzymes are also discussed here. Additionally, we provide suggestions about future directions in the study of enzymes in S. suis.

Download Full-text

Candidate proteomic biomarkers for three genogroups of the swine pathogen Streptococcus suis serotype 2

BMC Microbiology ◽

10.1186/s12866-015-0401-0 ◽

2015 ◽

Vol 15 (1) ◽

Cited By ~ 3

Author(s):

Christo Atanassov ◽

Laetitia Bonifait ◽

Marylise Perivier ◽

Marcelo Gottschalk ◽

Daniel Grenier

Keyword(s):

Streptococcus Suis ◽

Swine Pathogen ◽

Suis Serotype ◽

Streptococcus Suis Serotype 2

Download Full-text

Proteolytic Processing of the Mycoplasma hyopneumoniae Cilium Adhesin

Infection and Immunity ◽

10.1128/iai.72.5.2791-2802.2004 ◽

2004 ◽

Vol 72 (5) ◽

pp. 2791-2802 ◽

Cited By ~ 82

Author(s):

Steven P. Djordjevic ◽

Stuart J. Cordwell ◽

Michael A. Djordjevic ◽

Jody Wilton ◽

F. Chris Minion

Keyword(s):

Signal Sequence ◽

Surface Protein ◽

Mycoplasma Hyopneumoniae ◽

Proteolytic Processing ◽

Secreted Proteins ◽

Immunogold Electron Microscopy ◽

Swine Pathogen ◽

C Terminus ◽

J Proteins ◽

Ciliated Epithelial Cells

ABSTRACT Mycoplasma hyopneumoniae is an economically significant swine pathogen that colonizes the respiratory ciliated epithelial cells. Cilium adherence is mediated by P97, a surface protein containing a repeating element (R1) that is responsible for binding. Here, we show that the cilium adhesin is proteolytically processed on the surface. Proteomic analysis of strain J proteins identified cleavage products of 22, 28, 66, and 94 kDa. N-terminal sequencing showed that the 66- and 94-kDa proteins possessed identical N termini and that the 66-kDa variant was generated by cleavage of the 28-kDa product from the C terminus. The 22-kDa product represented the N-terminal 195 amino acids of the cilium adhesin preprotein, confirming that the hydrophobic leader signal sequence is not cleaved during translocation across the membrane. Comparative studies of M. hyopneumoniae strain 232 showed that the major cleavage products of the cilium adhesin are similar, although P22 and P28 appear to be processed further in strain 232. Immunoblotting studies using antisera raised against peptide sequences within P22 and P66/P94 indicate that processing is complex, with cleavage occurring at different frequencies within multiple sites, and is strain specific. Immunogold electron microscopy showed that fragments containing the cilium-binding site remained associated with the cell surface whereas cleavage products not containing the R1 element were located elsewhere. Not all secreted proteins undergo multiple cleavage, however, as evidenced by the analysis of the P102 gene product. The ability of M. hyopneumoniae to selectively cleave its secreted proteins provides this pathogen with a remarkable capacity to alter its surface architecture.

Download Full-text

Biochemical and cellular mechanisms regulatingAcanthamoeba castellaniiadherence to host cells

Parasitology ◽

10.1017/s0031182013001923 ◽

2013 ◽

Vol 141 (4) ◽

pp. 531-541 ◽

Cited By ~ 11

Author(s):

K. J. SOTO-ARREDONDO ◽

L. L. FLORES-VILLAVICENCIO ◽

J. J. SERRANO-LUNA ◽

M. SHIBAYAMA ◽

M. SABANERO-LÓPEZ

Keyword(s):

Neuronal Cells ◽

Acanthamoeba Castellanii ◽

Cell Types ◽

Infection Process ◽

Surface Proteins ◽

Host Cells ◽

Epifluorescence Microscopy ◽

Cellular Mechanisms ◽

Cutaneous Lesions ◽

Causative Agents

SUMMARYFree-living amoebae belonging to the genusAcanthamoebaare the causative agents of infections such as amoebic keratitis (AK), granulomatous amoebic encephalitis (GAE) and cutaneous lesions. The mechanisms involved in the establishment of infection are unknown. However, it is accepted that the initial phase of pathogenesis involves adherence to the host tissue. In this work, we analysed surface molecules with an affinity for epithelial and neuronal cells from the trophozoites ofAcanthamoeba castellanii. We also investigated the cellular mechanisms that govern the process of trophozoite adhesion to the host cells. We first used confocal and epifluorescence microscopy to examine the distribution of theA. castellaniiactin cytoskeleton during interaction with the host cells. The use of drugs, as cytochalasin B (CB) and latrunculin B (LB), revealed the participation of cytoskeletal filaments in the adhesion process. In addition, to identify the proteins and glycoproteins on the surface ofA. castellanii, the trophozoites were labelled with biotin and biotinylated lectins. The results revealed bands of surface proteins, some of which were glycoproteins with mannose andN-acetylglucosamine residues. Interaction assays of biotinylated amoebae proteins with epithelial and neuronal cells showed that some surface proteins had affinity for both cell types. The results of this study provide insight into the biochemical and cellular mechanisms of theAcanthamoebainfection process.

Download Full-text