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1.
J Infect Dis ; 227(9): 1050-1058, 2023 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-36588346

RESUMO

Mycoplasma penetrans is an emerging pathogen with a reduced genome. This bacterium has only previously been cultured from individuals with chronic immunodeficiencies. Here we report the characteristics of 4 M. penetrans isolates from the urine of immunocompetent males with nongonococcal urethritis, in comparison with strain HF-2 from an immunocompromised patient. Several features exhibited distinct differences between these isolates and HF-2. Unlike HF-2, all 4 were resistant to azithromycin. They exhibited greater sialic acid-dependent binding to erythrocytes, gliding motility speed, and H2O2 production than HF-2. All new isolates produced thinner capsules than HF-2. Invasiveness varied, with some isolates being more invasive than HF-2 and some less invasive. Cytotoxicity to HeLa cells was similar to HF-2, and all strains could clear extracellular traps produced by innate immune cells. We conclude that subtle differences among M. penetrans strains may be critical for this organism to establish an infection in an otherwise healthy individual.


Assuntos
Infecções por Mycoplasma , Mycoplasma penetrans , Uretrite , Masculino , Humanos , Uretrite/microbiologia , Células HeLa , Peróxido de Hidrogênio , Virulência , Infecções por Mycoplasma/microbiologia
2.
Clin Infect Dis ; 73(7): e1684-e1693, 2021 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-32750107

RESUMO

BACKGROUND: Nongonococcal urethritis (NGU) is a common syndrome with no known etiology in ≤50% of cases. We estimated associations between urethral bacteria and NGU in men who have sex with men (MSM) and men who have sex with women (MSW). METHODS: Urine was collected from NGU cases (129 MSM, 121 MSW) and controls (70 MSM, 114 MSW) attending a Seattle STD clinic. Cases had ≥5 polymorphonuclear leukocytes on Gram stain plus symptoms or discharge; controls had <5 PMNs, no symptoms, no discharge. NGU was considered idiopathic when Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Trichomonas vaginalis, adenovirus, and herpes simplex virus were absent. The urethral microbiota was characterized using 16S rRNA gene sequencing. Compositional lasso analysis was conducted to identify associations between bacterial taxa and NGU and to select bacteria for targeted qPCR. RESULTS: Among NGU cases, 45.2% were idiopathic. Based on compositional lasso analysis, we selected Haemophilus influenzae (HI) and Mycoplasma penetrans (MP) for targeted qPCR. Compared with 182 men without NGU, the 249 men with NGU were more likely to have HI (14% vs 2%) and MP (21% vs 1%) (both P ≤ .001). In stratified analyses, detection of HI was associated with NGU among MSM (12% vs 3%, P = .036) and MSW (17% vs 1%, P < .001), but MP was associated with NGU only among MSM (13% vs 1%, P = .004). Associations were stronger in men with idiopathic NGU. CONCLUSIONS: HI and MP are potential causes of male urethritis. MP was more often detected among MSM than MSW with urethritis.


Assuntos
Microbiota , Infecções por Mycoplasma , Mycoplasma penetrans , Minorias Sexuais e de Gênero , Uretrite , Chlamydia trachomatis , Feminino , Haemophilus influenzae , Homossexualidade Masculina , Humanos , Masculino , RNA Ribossômico 16S/genética , Comportamento Sexual
3.
Genes (Basel) ; 11(11)2020 11 07.
Artigo em Inglês | MEDLINE | ID: mdl-33171705

RESUMO

The structural organization and functionality of aminoacyl-tRNA synthetases have been expanded through polypeptide additions to their core aminoacylation domain. We have identified a novel domain appended to the methionyl-tRNA synthetase (MetRS) of the intracellular pathogen Mycoplasma penetrans. Sequence analysis of this N-terminal region suggests the appended domain is an aminotransferase, which we demonstrate here. The aminotransferase domain of MpMetRS is capable of generating methionine from its α-keto acid analog, 2-keto-4-methylthiobutyrate (KMTB). The methionine thus produced can be subsequently attached to cognate tRNAMet in the MpMetRS aminoacylation domain. Genomic erosion in the Mycoplasma species has impaired many canonical biosynthetic pathways, causing them to rely on their host for numerous metabolites. It is still unclear if this bifunctional MetRS is a key part of pathogen life cycle or is a neutral consequence of the reductive evolution experienced by Mycoplasma species.


Assuntos
Metionina tRNA Ligase/química , Metionina tRNA Ligase/metabolismo , Mycoplasma penetrans/genética , Sequência de Aminoácidos/genética , Aminoácidos/genética , Aminoacil-tRNA Sintetases/química , Aminoacil-tRNA Sintetases/metabolismo , Sítios de Ligação/genética , Metionina/análogos & derivados , Metionina/metabolismo , Domínios Proteicos/genética , RNA de Transferência/genética , Transaminases/genética , Transaminases/metabolismo
4.
BMC Infect Dis ; 20(1): 7, 2020 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-31900105

RESUMO

BACKGROUND: Mycoplasma sp. are well recognized as etiological agents of respiratory and sexually transmitted disease. Mycoplasma penetrans, a species of Mycoplasma sp., has been frequently detected in HIV-positive patients and associated with the progression of HIV-associated disease. To date, there is only a single case report describing M. penetrans as the causative agent of a severe respiratory tract infection in a HIV-negative patient. CASE PRESENTATION: In this report, we describe the case of M. penetrans bacteremia in a HIV-negative, 38-year-old, female, immunocompromised, solid organ transplant patient (combined kidney and pancreas transplantation in 2016), who was admitted to our hospital with anemic uterine bleeding and fever of 38.3 °C. Several hours before her admission at our university hospital, a latex bladder catheter was inserted into her uterus and she complained about fatigue, dizziness and ongoing vaginal bleeding. Laboratory examination showed severe anemia, but microbiological examination was inconspicuous (culture negative vaginal and cervical smears, negative urine culture). Bacterial blood cultures showed a growth signal after 4 h, but microscopic examination with Gram staining and subcultures on different agar media did not identify bacterial pathogens. To identify the bacterial cause of malignancy in the patient, metagenomic sequencing of the blood culture was performed that identified M. penetrans. CONCLUSION: Metagenomic sequencing identified M. penetrans in an immunosuppressed patient with culture-negative bacteremia. Clinicians should be aware of the opportunistic potential of M. penetrans that may cause severe infections in certain vulnerable patient populations and the limitations of culture and Gram staining for confirming the presence of fastidious bacterial pathogens like Mycoplasma spp.


Assuntos
Bacteriemia/diagnóstico , Hospedeiro Imunocomprometido , Metagenômica , Infecções por Mycoplasma/diagnóstico , Mycoplasma penetrans , Infecções Respiratórias/diagnóstico , Adulto , Bacteriemia/genética , Bacteriemia/microbiologia , Análise Mutacional de DNA/métodos , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/terapia , Feminino , Soronegatividade para HIV , Humanos , Transplante de Rim , Metagenoma , Metagenômica/métodos , Infecções por Mycoplasma/genética , Infecções por Mycoplasma/microbiologia , Mycoplasma penetrans/genética , Mycoplasma penetrans/isolamento & purificação , Transplante de Pâncreas , Infecções Respiratórias/genética , Infecções Respiratórias/microbiologia , Análise de Sequência de DNA
5.
Recent Pat Biotechnol ; 13(2): 124-136, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30569861

RESUMO

BACKGROUND: Arginine deiminase (ADI), an arginine catabolizing enzyme, is considered as an anti-tumor agent for the treatment of arginine auxotrophic cancers. However, some obstacles limit its clinical applications. OBJECTIVE: This review will summarize the clinical applications of ADI, from a brief history to its limitations, and will discuss the different ways to deal with the clinical limitations. METHOD: The structure analysis, cloning, expression, protein engineering and applications of arginine deiminase enzyme have been explained in this review. CONCLUSION: Recent patents on ADI are related to ADI engineering to increase its efficacy for clinical application. The intracellular delivery of ADI and combination therapy seem to be the future strategies in the treatment of arginine auxotrophic cancers. Applying ADIs with optimum features from different sources and or ADI engineering, are promising strategies to improve the clinical application of ADI.


Assuntos
Antineoplásicos/metabolismo , Arginina/metabolismo , Hidrolases/genética , Neoplasias/tratamento farmacológico , Engenharia de Proteínas/métodos , Antineoplásicos/uso terapêutico , Ensaios Clínicos como Assunto , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Humanos , Hidrolases/biossíntese , Hidrolases/uso terapêutico , Modelos Moleculares , Mycoplasma/química , Mycoplasma/enzimologia , Mycoplasma penetrans/química , Mycoplasma penetrans/enzimologia , Neoplasias/enzimologia , Neoplasias/patologia , Patentes como Assunto , Estrutura Secundária de Proteína , Pseudomonas aeruginosa/química , Pseudomonas aeruginosa/enzimologia , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
6.
PLoS One ; 13(5): e0197232, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29746549

RESUMO

The highly similar prokaryotic DNA (cytosine-5) methyltransferases (C5-MTases) M.MpeI and M.SssI share the specificity of eukaryotic C5-MTases (5'-CG), and can be useful research tools in the study of eukaryotic DNA methylation and epigenetic regulation. In an effort to improve the stability and solubility of complementing fragments of the two MTases, genes encoding circularly permuted (CP) variants of M.MpeI and M.SssI were created, and cloned in a plasmid vector downstream of an arabinose-inducible promoter. MTase activity of the CP variants was tested by digestion of the plasmids with methylation-sensitive restriction enzymes. Eleven of the fourteen M.MpeI permutants and six of the seven M.SssI permutants had detectable MTase activity as indicated by the full or partial protection of the plasmid carrying the cpMTase gene. Permutants cp62M.MpeI and cp58M.SssI, in which the new N-termini are located between conserved motifs II and III, had by far the highest activity. The activity of cp62M.MpeI was comparable to the activity of wild-type M.MpeI. Based on the location of the split sites, the permutants possessing MTase activity can be classified in ten types. Although most permutation sites were designed to fall outside of conserved motifs, and the MTase activity of the permutants measured in cell extracts was in most cases substantially lower than that of the wild-type enzyme, the high proportion of circular permutation topologies compatible with MTase activity is remarkable, and is a new evidence for the structural plasticity of C5-MTases. A computer search of the REBASE database identified putative C5-MTases with CP arrangement. Interestingly, all natural circularly permuted C5-MTases appear to represent only one of the ten types of permutation topology created in this work.


Assuntos
Proteínas de Bactérias/química , DNA (Citosina-5-)-Metiltransferases/química , Bases de Dados de Proteínas , Mycoplasma penetrans/enzimologia , Spiroplasma/enzimologia , Proteínas de Bactérias/genética , DNA (Citosina-5-)-Metiltransferases/genética , Mycoplasma penetrans/genética , Spiroplasma/genética , Especificidade por Substrato
7.
J Fish Dis ; 41(4): 569-579, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29023774

RESUMO

Intestinal neoplasms are common in zebrafish (Danio rerio) research facilities. These tumours are most often seen in older fish and are classified as small cell carcinomas or adenocarcinomas. Affected fish populations always contain subpopulations with preneoplastic lesions, characterized by epithelial hyperplasia or inflammation. Previous observations indicated that these tumours are unlikely caused by diet, water quality or genetic background, suggesting an infectious aetiology. We performed five transmission experiments by exposure of naïve fish to affected donor fish by cohabitation or exposure to tank effluent water. Intestinal lesions were observed in recipient fish in all exposure groups, including transmissions from previous recipient fish, and moribund fish exhibited a higher prevalence of neoplasms. We found a single 16S rRNA sequence, most similar to Mycoplasma penetrans, to be highly enriched in the donors and exposed recipients compared to unexposed control fish. We further tracked the presence of the Mycoplasma sp. using a targeted PCR test on individual dissected intestines or faeces or tank faeces. Original donor and exposed fish populations were positive for Mycoplasma, while corresponding unexposed control fish were negative. This study indicates an infectious aetiology for these transmissible tumours of zebrafish and suggests a possible candidate agent of a Mycoplasma species.


Assuntos
Doenças dos Peixes/transmissão , Neoplasias Intestinais , Infecções por Mycoplasma/transmissão , Mycoplasma penetrans/isolamento & purificação , Mycoplasma penetrans/fisiologia , Peixe-Zebra , Adenocarcinoma/microbiologia , Animais , Carcinoma de Células Pequenas/microbiologia , Doenças dos Peixes/microbiologia , Neoplasias Intestinais/microbiologia , Infecções por Mycoplasma/microbiologia , Mycoplasma penetrans/genética , RNA Bacteriano/genética , RNA Ribossômico 16S/genética
8.
PLoS One ; 12(7): e0180514, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28692662

RESUMO

Mycoplasma infection has been reported in immunocompromised cancer patients; nevertheless, it is not clear if persistent Mycoplasma infection could facilitate the proliferation of cancer cells in immunocompromised organisms. The aim of this study was to examine the relationship between persistent Mycoplasma infection and malignant transformation in an immunodeficient host model. Immunodeficient mouse model was established using cyclophosphamide and mice gastric mucosal cells were infected with Mycoplasma penetrans (Mpe). After 18 weeks, mice were sacrificed and gastric mucosal Mpe infected cells were identified by fluorescence in situ hybridization (FISH). Moreover, pathological and ultrastructural changes in mice gastric mucosa were evaluated and the expression of multiple proto-oncogenes was examined by Western blot. Our data show that Mpe infection was detected in the blood of immunodeficient mice and Mpe persistent infection in mice gastric mucosa was confirmed by FISH. There were pathological and ultrastructural malignant transformation occurred in the gastric mucosa of infected mice compared to control mice. Mpe infected mice showed lower expression of p53 and p21 and higher H-ras expression compared to the control group. Moreover, expression of NF-κB p65 subunit increased in Mpe infected mice, similar to the TNF-α expression. Bax expression in gastric mucosa of Mpe infected mice was lower while Bcl-2 expression was higher than in the uninfected control group. Collectively these data demonstrate that persistent Mpe infection is associated with aberrant expression of multiple proto-oncogenes in gastric mucosa of immunodeficient mice which potentially facilitate the malignant transformation.


Assuntos
Transformação Celular Neoplásica/patologia , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/patologia , Mycoplasma penetrans/fisiologia , Animais , Apoptose , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Feminino , Mucosa Gástrica/ultraestrutura , Camundongos Endogâmicos C57BL , Camundongos SCID , Infecções por Mycoplasma/diagnóstico , Mycoplasma penetrans/ultraestrutura , NF-kappa B/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo , Proteínas ras/metabolismo
9.
J Bacteriol ; 199(12)2017 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-28373274

RESUMO

Although mycoplasmas have small genomes, many of them, including the HIV-associated opportunist Mycoplasma penetrans, construct a polar attachment organelle (AO) that is used for both adherence to host cells and gliding motility. However, the irregular phylogenetic distribution of similar structures within the mycoplasmas, as well as compositional and ultrastructural differences among these AOs, suggests that AOs have arisen several times through convergent evolution. We investigated the ultrastructure and protein composition of the cytoskeleton-like material of the M. penetrans AO with several forms of microscopy and biochemical analysis, to determine whether the M. penetrans AO was constructed at the molecular level on principles similar to those of other mycoplasmas, such as Mycoplasma pneumoniae and Mycoplasma mobile We found that the M. penetrans AO interior was generally dissimilar from that of other mycoplasmas, in that it exhibited considerable heterogeneity in size and shape, suggesting a gel-like nature. In contrast, several of the 12 potential protein components identified by mass spectrometry of M. penetrans detergent-insoluble proteins shared certain distinctive biochemical characteristics with M. pneumoniae AO proteins, although not with M. mobile proteins. We conclude that convergence between M. penetrans and M. pneumoniae AOs extends to the molecular level, leading to the possibility that the less organized material in both M. pneumoniae and M. penetrans is the substance principally responsible for the organization and function of the AO.IMPORTANCEMycoplasma penetrans is a bacterium that infects HIV-positive patients and may contribute to the progression of AIDS. It attaches to host cells through a structure called an AO, but it is not clear how it builds this structure. Our research is significant not only because it identifies the novel protein components that make up the material within the AO that give it its structure but also because we find that the M. penetrans AO is organized unlike AOs from other mycoplasmas, suggesting that similar structures have evolved multiple times. From this work, we derive some basic principles by which mycoplasmas, and potentially all organisms, build structures at the subcellular level.


Assuntos
Estruturas Bacterianas/química , Estruturas Bacterianas/ultraestrutura , Mycoplasma penetrans/química , Mycoplasma penetrans/ultraestrutura , Organelas/química , Organelas/ultraestrutura , Evolução Biológica , Espectrometria de Massas , Mycoplasma pneumoniae/química , Mycoplasma pneumoniae/fisiologia , Mycoplasma pneumoniae/ultraestrutura
10.
Appl Microbiol Biotechnol ; 100(11): 4747-60, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27087524

RESUMO

Arginine deiminase (ADI) is an important arginine-degrading enzyme with wide applications, in particular as an anti-cancer agent for the therapy of arginine-auxotrophic tumors. In recent years, novel ADIs with excellent properties have been identified from various organisms, and crystal structures of ADI were investigated. To satisfy the requirements of potential therapeutic applications, protein engineering has been performed to improve the activity and properties of ADIs. In this mini-review, we systematically summarized the latest progress on identification and crystal structure of ADIs, and protein engineering strategies for improved enzymatic properties, such as pH optimum, K m and k cat values, and thermostability. We also outlined the PEGylation of ADI for improved circulating half-life and immunogenicity, as well as their performance in clinical trials. Finally, perspectives on extracellular secretion and property improvement of ADI were discussed.


Assuntos
Antineoplásicos/química , Hidrolases/química , Engenharia de Proteínas , Animais , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Ensaios Clínicos como Assunto , Modelos Animais de Doenças , Sinergismo Farmacológico , Humanos , Concentração de Íons de Hidrogênio , Hidrolases/farmacologia , Mycoplasma/classificação , Mycoplasma/enzimologia , Mycoplasma penetrans/enzimologia , Neoplasias/tratamento farmacológico , Conformação Proteica , Pseudomonas aeruginosa/enzimologia
11.
Epidemiol Infect ; 143(15): 3327-34, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25792346

RESUMO

Mycoplasma infections are most frequently associated with disease in the urogenital or respiratory tracts and, in most cases, mycoplasmas infect the host persistently. In HIV-infected individuals the prevalence and role of genital mycoplasmas has not been well studied. To investigate the six species of Mycoplasma and the risk factors for infection in Jiangsu province, first-void urine and venous blood samples were collected and epidemiological questionnaires were administered after informed consent. A total of 1541 HIV/AIDS patients were recruited in this study. The overall infection rates of six Mycoplasma species were: Ureaplasma urealyticum (26·7%), Mycoplasma hominis (25·3%), M. fermentans (5·1%), M. genitalium (20·1%), M. penetrans (1·6%) and M. pirum (15·4%). The Mycoplasma infection rate in the unmarried group was lower than that of the married, divorced and widowed groups [adjusted odds ratio (aOR) 1·432, 95% confidence interval (CI) 1·077-1·904, P < 0·05]. The patients who refused highly active antiretroviral therapy (HAART) had a much higher risk of Mucoplasma infection (aOR 1·357, 95% CI 1·097-1·679, P < 0·05). Otherwise, a high CD4+ T cell count was a protective factor against Mycoplasma infection (aOR 0·576, 95% CI 0·460-0·719, P < 0·05). Further research will be required to confirm a causal relationship and to identify risk factors for Mycoplasma infection in HIV/AIDS populations.


Assuntos
Terapia Antirretroviral de Alta Atividade/estatística & dados numéricos , Infecções por HIV/epidemiologia , Estado Civil/estatística & dados numéricos , Infecções por Mycoplasma/epidemiologia , Infecções por Ureaplasma/epidemiologia , Adolescente , Adulto , Idoso , China/epidemiologia , Coinfecção/epidemiologia , Estudos Transversais , Infecções por HIV/tratamento farmacológico , HIV-1 , Humanos , Masculino , Pessoa de Meia-Idade , Mycoplasma/genética , Mycoplasma/isolamento & purificação , Infecções por Mycoplasma/microbiologia , Mycoplasma fermentans/genética , Mycoplasma fermentans/isolamento & purificação , Mycoplasma genitalium/genética , Mycoplasma genitalium/isolamento & purificação , Mycoplasma hominis/genética , Mycoplasma hominis/isolamento & purificação , Mycoplasma penetrans/genética , Mycoplasma penetrans/isolamento & purificação , Reação em Cadeia da Polimerase , Prevalência , Fatores de Risco , Ureaplasma urealyticum/genética , Ureaplasma urealyticum/isolamento & purificação , Adulto Jovem
12.
FEMS Microbiol Lett ; 338(1): 39-45, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23066969

RESUMO

Mycoplasma penetrans, a potential human pathogen found mainly in HIV-infected individuals, uses a tip structure for both adherence and gliding motility. To improve our understanding of the molecular mechanism of M. penetrans gliding motility, we used chemical inhibitors of energy sources associated with motility of other organisms to determine which of these is used by M. penetrans and also tested whether gliding speed responded to temperature and pH. Mycoplasma penetrans gliding motility was not eliminated in the presence of a proton motive force inhibitor, a sodium motive force inhibitor, or an agent that depletes cellular ATP. At near-neutral pH, gliding speed increased as temperature increased. The absence of a clear chemical energy source for gliding motility and a positive correlation between speed and temperature suggest that energy derived from heat provides the major source of power for the gliding motor of M. penetrans.


Assuntos
Metabolismo Energético , Temperatura Alta , Mycoplasma penetrans/fisiologia , Fenômenos Fisiológicos Bacterianos , Humanos , Concentração de Íons de Hidrogênio , Microscopia de Fluorescência , Microscopia de Contraste de Fase , Mycoplasma penetrans/metabolismo , Temperatura , Imagem com Lapso de Tempo
13.
J Nephrol ; 26(3): 470-5, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-22782329

RESUMO

AIM: A new animal model of immunoglobulin A nephropathy (IgAN) was made by infecting mice with Mycoplasma penetrans (Mpe). To examine the pathogenesis of IgAN induced by Mpe infection, tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and nuclear factor-kB (NF-kB) protein levels were compared among study groups. METHODS: To make an experimental IgAN animal model, mice were infected with Mpe, SP-4 medium or phosphate-buffered saline (PBS) via the urinary tract. To compare changes in the classical IgAN model, TNF-alpha and IL-6 RNA expression levels were measured using RT-PCR, and NF-kB protein was measured using EMSA. RESULTS: By producing a urinary tract infection with Mpe, we developed a new animal model of IgAN with a 100% success rate. There was no difference with the classical animal model. We also observed IgG deposition in 66.67% of the Mpe-infection group. Glomerular cell and mesangial matrix proliferation was greater in the Mpe-infection group than in the control groups (p<0.05). In the Mpe-infection and classical groups, TNF-alpha and IL-6 expression levels were much higher than in the control groups (p<0.01). NF-kB expression was much higher in the Mpe-infection group (p<0.05). CONCLUSIONS: We made a new IgAN animal model that will offer a new direction for IgAN research. The activation of inflammation factors was associated with the Mpe induction of IgAN.


Assuntos
Modelos Animais de Doenças , Glomerulonefrite por IGA/microbiologia , Infecções por Mycoplasma/complicações , Mycoplasma penetrans , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C
14.
Proc Natl Acad Sci U S A ; 110(1): 105-10, 2013 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-23248272

RESUMO

Cytosine methylation promotes deamination. In eukaryotes, CpG methylation is thought to account for CpG underrepresentation. Whether scarcity of CpGs in prokaryotic genomes is diagnostic for methylation is not clear. Here, we report that Mycoplasms tend to be CpG depleted and to harbor a family of constitutively expressed or phase variable CpG-specific DNA methyltransferases. The very CpG poor Mycoplasma penetrans and its constitutively active CpG-specific methyltransferase M.MpeI were chosen for further characterization. Genome-wide sequencing of bisulfite-converted DNA indicated that M.MpeI methylated CpG target sites both in vivo and in vitro in a locus-nonselective manner. A crystal structure of M.MpeI with DNA at 2.15-Å resolution showed that the substrate base was flipped and that its place in the DNA stack was taken by a glutamine residue. A phenylalanine residue was intercalated into the "weak" CpG step of the nonsubstrate strand, indicating mechanistic similarities in the recognition of the short CpG target sequence by prokaryotic and eukaryotic DNA methyltransferases.


Assuntos
Ilhas de CpG/genética , DNA (Citosina-5-)-Metiltransferases/química , DNA (Citosina-5-)-Metiltransferases/metabolismo , DNA/química , Modelos Moleculares , Mycoplasma penetrans/enzimologia , Mycoplasma penetrans/genética , Sequência de Bases , Clonagem Molecular , Cristalização , DNA/metabolismo , Desaminação , Sequenciamento de Nucleotídeos em Larga Escala , Dados de Sequência Molecular , Especificidade por Substrato
15.
PLoS One ; 7(10): e47886, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23082227

RESUMO

The metabolism of arginine towards ATP synthesis has been considered a major source of energy for microorganisms such as Mycoplasma penetrans in anaerobic conditions. Additionally, this pathway has also been implicated in pathogenic and virulence mechanism of certain microorganisms, i.e. protection from acidic stress during infection. In this work we present the crystal structures of the three enzymes composing the gene cluster of the arginine deiminase pathway from M. penetrans: arginine deiminase (ADI), ornithine carbamoyltransferase (OTC) and carbamate kinase (CK). The arginine deiminase (ADI) structure has been refined to 2.3 Å resolution in its apo-form, displaying an "open" conformation of the active site of the enzyme in comparison to previous complex structures with substrate intermediates. The active site pocket of ADI is empty, with some of the catalytic and binding residues far from their active positions, suggesting major conformational changes upon substrate binding. Ornithine carbamoyltransferase (OTC) has been refined in two crystal forms at 2.5 Å and 2.6 Å resolution, respectively, both displaying an identical dodecameric structure with a 23-point symmetry. The dodecameric structure of OTC represents the highest level of organization in this protein family and in M.penetrans it is constituted by a novel interface between the four catalytic homotrimers. Carbamate kinase (CK) has been refined to 2.5 Å resolution and its structure is characterized by the presence of two ion sulfates in the active site, one in the carbamoyl phosphate binding site and the other in the ß-phosphate ADP binding pocket of the enzyme. The CK structure also shows variations in some of the elements that regulate the catalytic activity of the enzyme. The relatively low number of metabolic pathways and the relevance in human pathogenesis of Mycoplasma penetrans places the arginine deiminase pathway enzymes as potential targets to design specific inhibitors against this human parasite.


Assuntos
Hidrolases/química , Redes e Vias Metabólicas , Mycoplasma penetrans/enzimologia , Ornitina Carbamoiltransferase/química , Fosfotransferases (Aceptor do Grupo Carboxila)/química , Cristalografia por Raios X , Humanos , Modelos Moleculares , Multimerização Proteica , Especificidade por Substrato
16.
Int J STD AIDS ; 23(8): e7-10, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22930310

RESUMO

This study employed culture and polymerase chain reaction (PCR) to examine the prevalence of Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium, Mycoplasma fermentans, Mycoplasma penetrans and Mycoplasma pirum in 210 HIV/AIDS patients, 455 sexually transmitted infection (STI) clinic attendees and 245 healthy volunteers from first-void urine specimens for men and endocervical swabs for women. U. urealyticum and M. hominis were detected in 107 (51.0%) and 69 (32.9%) patients in the HIV/AIDS group. At least one of the other four organisms was detected in 34 (16.2%) HIV/AIDS patients, 29 (6.4%) STI clinic attendees and six (2.5%) healthy volunteers. This study showed that U. urealyticum, M. hominis and M. fermentans were significantly more prevalent in HIV/AIDS patients, as were other mycoplasmas. Our results suggest a possible role for co-infection.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Mycoplasmataceae , Infecções por Mycoplasmatales/epidemiologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mycoplasma , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Mycoplasma fermentans , Mycoplasma genitalium , Mycoplasma hominis , Mycoplasma penetrans , Infecções por Mycoplasmatales/microbiologia , Reação em Cadeia da Polimerase , Prevalência , Infecções por Ureaplasma/epidemiologia , Infecções por Ureaplasma/microbiologia , Ureaplasma urealyticum , Adulto Jovem
17.
J Bacteriol ; 194(11): 2877-83, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22447904

RESUMO

Within the genus Mycoplasma are species whose cells have terminal organelles, polarized structures associated with cytadherence and gliding motility. Mycoplasma penetrans, found mostly in HIV-infected patients, and Mycoplasma iowae, an economically significant poultry pathogen, are members of the Mycoplasma muris phylogenetic cluster. Both species have terminal organelles that interact with host cells, yet the structures in these species, or any in the M. muris cluster, remain uncharacterized. Time-lapse microcinematography of two strains of M. penetrans, GTU-54-6A1 and HF-2, and two serovars of M. iowae, K and N, show that the terminal organelles of both species play a role in gliding motility, with differences in speed within and between the two species. The strains and serovars also differed in their hemadsorption abilities that positively correlated with differences in motility speeds. No morphological differences were observed between M. penetrans and M. iowae by scanning electron microscopy (SEM). SEM and light microscopy of M. penetrans and M. iowae showed the presence of membranous filaments connecting pairs of dividing cells. Breaking of this filament during cell division was observed for M. penetrans by microcinematography, and this suggests a role for motility during division. The Triton X-100-insoluble fractions of M. penetrans and M. iowae consisted of similar structures that were unique compared to those identified in other mycoplasma species. Like other polarized mycoplasmas, M. penetrans and M. iowae have terminal organelles with cytadherence and gliding functions. The difference in function and morphology of the terminal organelles suggests that mycoplasmas have evolved terminal organelles independently of one another.


Assuntos
Mycoplasma iowae/fisiologia , Mycoplasma penetrans/fisiologia , Organelas/fisiologia , Microscopia Eletrônica de Varredura , Mycoplasma iowae/citologia , Mycoplasma iowae/crescimento & desenvolvimento , Mycoplasma iowae/ultraestrutura , Mycoplasma penetrans/citologia , Mycoplasma penetrans/crescimento & desenvolvimento , Mycoplasma penetrans/ultraestrutura , Organelas/ultraestrutura , Imagem com Lapso de Tempo
18.
Infect Immun ; 77(10): 4362-70, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19651868

RESUMO

Mycoplasma penetrans is a urogenital tract pathogen implicated in the deterioration of the immune system in human immunodeficiency virus-infected AIDS patients. Here, we describe a 78-kDa protein from M. penetrans, designated MYPE9110, that exhibits sequence similarity to known ADP-ribosyltransferases (ADPRTs) such as Bordetella pertussis pertussis toxin and Mycoplasma pneumoniae community-acquired respiratory distress syndrome toxin. MYPE9110 possesses key amino acid residues found in all ADPRTs that are essential for ADPRT activity. Several mammalian cell proteins are ADP-ribosylated by MYPE9110, and the full-length recombinant protein exhibits a strong auto-ADP-ribosylating activity. In the absence of target proteins, MYPE9110 demonstrates a NAD-glycohydrolase activity by hydrolyzing NAD. Furthermore, this toxin elicits cytopathology in HeLa cells by inducing cytoplasmic vacuolization in the presence of ammonium chloride. The deletion of the C-terminal region of MYPE9110 significantly diminishes its binding to host cells while still exhibiting an ADPRT activity, suggesting that MYPE9110 is a member of the family of A-B ADPRT toxins.


Assuntos
ADP Ribose Transferases/genética , ADP Ribose Transferases/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Mycoplasma penetrans/enzimologia , Bordetella pertussis/genética , Ordem dos Genes , Genes Bacterianos , Células HeLa , Humanos , Mycoplasma pneumoniae/genética , Homologia de Sequência de Aminoácidos
19.
Microbiology (Reading) ; 155(Pt 4): 1241-1249, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19332825

RESUMO

Mycoplasma penetrans has the ability to change its surface lipoprotein profiles frequently. The P35 family lipoproteins encoded by the mpl genes are key players in this profile variation. The M. penetrans HF-2 genome has 38 mpl genes that form three gene clusters. Most of these mpl genes have an invertible promoter sequence that is responsible for the ON/OFF switching of individual mpl gene expression. Here, we identified the recombinase that catalyses inversions of the mpl gene promoters. We focused on two open reading frames of the M. penetrans HF-2 genome, namely MYPE2900 and MYPE8180, which show significant homology to the tyrosine site-specific recombinase (Tsr) family proteins. Since genetic tools for M. penetrans are still not developed, we cloned the MYPE2900 and MYPE8180 genes and expressed them in Mycoplasma pneumoniae and Escherichia coli. The promoter regions of the mpl genes [p35 (MYPE6810) or p42 (MYPE6630) genes] were also introduced into M. pneumoniae and E. coli cells expressing MYPE2900 or MYPE8180. Inversion of these promoters occurred in the presence of the MYPE2900 gene but not in the presence of the MYPE8180 gene, indicating that the MYPE2900 gene product is the recombinase that catalyses mpl gene promoter inversions. We used a PCR-based method to detect mpl promoter inversion. This method also enabled us to detect inversions of 10 mpl gene promoters in M. penetrans HF-2 cells. All these promoter inversions occurred at the 12 bp inverted repeat (IR) sequences flanking the promoter sequence. The consensus sequence of these IRs was proposed as TAAYNNNDATTA (Y=C or T; D=A, G or T).


Assuntos
Inversão Cromossômica , DNA Nucleotidiltransferases/metabolismo , Lipoproteínas/genética , Mycoplasma penetrans/enzimologia , Regiões Promotoras Genéticas , Proteínas de Bactérias/genética , Sequência de Bases , Clonagem Molecular , DNA Nucleotidiltransferases/genética , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Humanos , Sequências Repetidas Invertidas/genética , Dados de Sequência Molecular , Família Multigênica , Mycoplasma penetrans/genética , Mycoplasma pneumoniae/genética , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas/genética
20.
Curr Microbiol ; 58(4): 360-5, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19189179

RESUMO

The current study shows that Mycoplasma penetrans strain GTU binds human serum immunoglobulin A (IgA) and secretory IgA but not IgG. Binding of IgA was associated almost exclusively with the lipoprotein fraction obtained by Triton X-114 fractionation of isolated M. penetrans membranes. Western immunoblot analysis of isolated membranes of M. penetrans strain GTU and of the Triton X-114 fraction showed that the major IgA-binding component was a lipoprotein with a molecular mass of 38 kDa, one of the major lipoproteins of this organism. The authors suggest that the high IgA-binding capacity of M. penetrans strain GTU may serve as a defense mechanism, conferring on this microorganism the ability to evade clearance mediated by specific IgA antibodies.


Assuntos
Antígenos de Bactérias/imunologia , Antígenos de Superfície/imunologia , Imunoglobulina A Secretora/imunologia , Imunoglobulina A/imunologia , Infecções por Mycoplasma/imunologia , Mycoplasma penetrans/imunologia , Adulto , Idoso , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Proteínas da Membrana Bacteriana Externa/metabolismo , Fracionamento Celular , Humanos , Imunidade Inata , Lactente , Lipoproteínas/química , Lipoproteínas/isolamento & purificação , Lipoproteínas/metabolismo , Peso Molecular , Mycoplasma penetrans/química , Octoxinol , Polietilenoglicóis
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