Assuntos
Células Epiteliais , Sindecana-1 , Fatores de Virulência , Humanos , Fatores de Virulência/metabolismo , Fatores de Virulência/genética , Células Epiteliais/microbiologia , Células Epiteliais/metabolismo , Sindecana-1/metabolismo , Sindecana-1/genética , Bacillus anthracis/metabolismo , Bacillus anthracis/patogenicidade , Bacillus anthracis/genética , Antraz/microbiologiaRESUMO
BACKGROUND: It has been recently reported that major pathogens Staphylococcus aureus and Pseudomonas aeruginosa accelerate a normal process of cell surface syndecan-1 (Synd1) ectodomain shedding as a mechanism of host damage due to the production of shedding-inducing virulence factors. We tested if acceleration of Synd1 shedding takes place in vitro upon treatment of epithelial cells with B. anthracis hemolysins, as well as in vivo during anthrax infection in mice. RESULTS: The isolated anthrax hemolytic proteins AnlB (sphingomyelinase) and AnlO (cholesterol-binding pore-forming factor), as well as ClnA (B. cereus homolog of B. anthracis phosphatidyl choline-preferring phospholipase C) cause accelerated shedding of Synd1 and E-cadherin from epithelial cells and compromise epithelial barrier integrity within a few hours. In comparison with hemolysins in a similar range of concentrations, anthrax lethal toxin (LT) also accelerates shedding albeit at slower rate. Individual components of LT, lethal factor and protective antigen are inactive with regard to shedding. Inhibition experiments favor a hypothesis that activities of tested bacterial shedding inducers converge on the stimulation of cytoplasmic tyrosine kinases of the Syk family, ultimately leading to activation of cellular sheddase. Both LT and AnlO modulate ERK1/2 and p38 MAPK signaling pathways, while JNK pathway seems to be irrelevant to accelerated shedding. Accelerated shedding of Synd1 also takes place in DBA/2 mice challenged with Bacillus anthracis (Sterne) spores. Elevated levels of shed ectodomain are readily detectable in circulation after 24 h. CONCLUSION: The concerted acceleration of shedding by several virulence factors could represent a new pathogenic mechanism contributing to disruption of epithelial or endothelial integrity, hemorrhage, edema and abnormal cell signaling during anthrax infection.
Assuntos
Bacillus anthracis/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Sindecana-1/metabolismo , Fatores de Virulência/metabolismo , Animais , Antígenos de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Caderinas/metabolismo , Linhagem Celular , Proteínas Hemolisinas/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Camundongos , Camundongos Endogâmicos DBA , Esfingomielina Fosfodiesterase/metabolismo , Fosfolipases Tipo C/metabolismoRESUMO
In the present study, we describe the human and mouse RFP2 gene structure, multiple RFP2 mRNA isoforms in the two species that have different 5' UTRs and a human-specific antisense transcript RFP2OS. Since the human RFP2 5' UTR is not conserved in mouse, these findings might indicate a different regulation of RFP2 in the two species. The predicted human and mouse RFP2 proteins are shown to contain a tripartite RING finger-B-box-coiled-coil domain (RBCC), also known as a TRIM domain, and therefore belong to a subgroup of RING finger proteins that are often involved in developmental and tumorigenic processes. Because homozygous deletions of chromosomal region 13q14.3 are found in a number of malignancies, including chronic lymphocytic leukemia (CLL) and multiple myeloma (MM), we suggest that RFP2 might be involved in tumor development. This study provides necessary information for evaluation of the role of RFP2 in malignant transformation and other biological processes.
Assuntos
Proteínas de Ligação a DNA/genética , RNA Antissenso/genética , RNA Mensageiro/genética , Proteínas Supressoras de Tumor/genética , Processamento Alternativo , Sequência de Aminoácidos , Animais , Mapeamento Cromossômico , Cromossomos Humanos Par 13/genética , Clonagem Molecular , DNA/química , DNA/genética , Éxons , Feminino , Expressão Gênica , Genes/genética , Humanos , Íntrons , Masculino , Camundongos , Dados de Sequência Molecular , Regiões Promotoras Genéticas/genética , RNA Antissenso/metabolismo , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Transcrição GênicaRESUMO
We have cloned the genes PANX1, PANX2 and PANX3, encoding putative gap junction proteins homologous to invertebrate innexins, which constitute a new family of mammalian proteins called pannexins. Phylogenetic analysis revealed that pannexins are highly conserved in worms, mollusks, insects and mammals, pointing to their important function. Both innexins and pannexins are predicted to have four transmembrane regions, two extracellular loops, one intracellular loop and intracellular N and C termini. Both the human and mouse genomes contain three pannexin-encoding genes. Mammalian pannexins PANX1 and PANX3 are closely related, with PANX2 more distant. The human and mouse pannexin-1 mRNAs are ubiquitously, although disproportionately, expressed in normal tissues. Human PANX2 is a brain-specific gene; its mouse orthologue, Panx2, is also expressed in certain cell types in developing brain. In silico evaluation of Panx3 expression predicts gene expression in osteoblasts and synovial fibroblasts. The apparent conservation of pannexins between species merits further investigation.