GroEL-like particles of Thermus aquaticus are homo-oligomeric complexes of two stacked seven member rings, sedimenting in the gradient at 20S. The apparent molecular mass of the native particles is 820,000 (+/- 30,000). The protein complex is composed with one polypeptide of M(r) 59,000. Immunoblotting results and N-terminal amino acid analysis indicate that the complex is significantly related to the chaperonins. No proteolytic activity was identified in the purified GroEL-like particles. In the presence of Mg2+ and K+ the complex exhibits temperature dependent ATPase activity. Under optimum temperature (75 degrees C) GroEL is stable and hydrolyze ATP with a specific activity of 0.47 mumol min-1 mg-1.

• MeSH
• Adenosine Triphosphatases isolation & purification   metabolism   MeSH
• Bacillus megaterium MeSH
• Bacterial Proteins chemistry   isolation & purification   metabolism   ultrastructure   MeSH
• Chaperonin 60 MeSH
• Chromatography, Ion Exchange MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Microscopy, Electron MeSH
• Endopeptidases metabolism   MeSH
• Kinetics MeSH
• Molecular Sequence Data MeSH
• Heat-Shock Proteins chemistry   isolation & purification   ultrastructure   MeSH
• Amino Acid Sequence MeSH
• Sequence Homology, Amino Acid MeSH
• Thermodynamics MeSH
• Thermus metabolism   MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• Adenosine Triphosphatases MeSH
• Bacterial Proteins MeSH
• Chaperonin 60 MeSH
• Endopeptidases MeSH
• Heat-Shock Proteins MeSH

Borrelia burgdorferi sensu lato (s.l.) is the etiological agent of Lyme disease, transmitted by ticks of the genus Ixodes Latreille. Diagnosis of Lyme disease in humans is often difficult and a detailed knowledge of the circulation of B. burgdorferi s.l. in tick hosts is therefore fundamental to support clinical procedures. Here we developed a molecular approach for the detection of B. burgdorferi s.l. in North Italian Ixodes ricinus (Linnaeus). The method is based on the amplification of a fragment of the groEL gene, which encodes a heat-shock protein highly conserved among B. burgdorferi s.l. species. The tool was applied in both qualitative and Real-time PCR approaches testing ticks collected in a North Italian area. The obtained results suggest that this new molecular tool could represent a sensitive and specific method for epidemiological studies aimed at defining the distribution of B. burgdorferi s.l. in I. ricinus and, consequently, the exposure risk for humans.

• Keywords
• Ixodes ricinus, Lyme diseases, Real-time PCR., detection, qualitative PCR,
• MeSH
• Bacterial Proteins analysis   MeSH
• Borrelia burgdorferi Group genetics   isolation & purification   MeSH
• Chaperonin 60 analysis   MeSH
• Ixodes growth & development   microbiology   MeSH
• Real-Time Polymerase Chain Reaction methods   MeSH
• Nymph growth & development   microbiology   MeSH
• Sequence Analysis, Protein MeSH
• Sequence Alignment MeSH
• Sensitivity and Specificity MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Italy MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Chaperonin 60 MeSH

GroEL or symbionin synthesized by the endosymbionts of whitefly (Bemisia tabaci)/ aphids play a cardinal role in the persistent, circulative transmission of plant viruses by binding to viral coat protein/ read-through protein. Allium sativum leaf agglutinin (ASAL), a Galanthus nivalis agglutinin (GNA)- related mannose-binding lectin from garlic leaf has been reported as a potent controlling agent against hemipteran insects including whitefly and aphids. GroEL related chaperonin- symbionin was previously identified as a receptor of ASAL by the present group in the brush border membrane vesicle (BBMV) of mustard aphid. In the present study similar GroEL receptor of ASAL has been identified through LC-MS/MS in the BBMV of B. tabaci which serves as a vector for several plant viruses including tomato leaf curl New Delhi virus (ToLCNDV). Ligand blot analysis of ASAL-fed B. tabaci showed that when GroEL is pre-occupied by ASAL, it completely blocks its further binding to ToLCNDV coat protein (ToLCNDV-CP). Prior feeding of ASAL hindered the co-localization of ToLCNDV-CP and GroEL in the midgut of B. tabaci. Immunoprecipitation followed by western blot with ASAL-fed B. tabaci yielded similar result. Moreover, ASAL feeding inhibited viral transmission by B. tabaci. Together, these results confirmed that the interaction of ASAL with GroEL interferes with the binding of ToLCNDV-CP and inhibits further B. tabaci mediated viral transmission.

• Keywords
• Allium sativum leaf agglutinin (ASAL), Bemisia tabaci, Endosymbiont GroEL, Persistent-circulative virus transmission, Tomato leaf curl New Delhi virus (ToLCNDV), Viral coat protein,
• MeSH
• Agglutinins MeSH
• Begomovirus * genetics   MeSH
• Garlic * MeSH
• Chromatography, Liquid MeSH
• Hemiptera * MeSH
• Lectins MeSH
• Aphids * MeSH
• Plant Diseases MeSH
• Tandem Mass Spectrometry MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Agglutinins MeSH
• Lectins MeSH

Cells of Bacillus megaterium 27 were challenged by a 30-min heat shock at 45 degrees C during various sporulation stages and then shifted back to a temperature permissive for sporulation (27 degrees C), at which they developed spores. Heat shock applied at 120 min after the end of the exponential phase induced synthesis of heat shock proteins (HSPs) in the sporangia and delayed the inactivation of spores at 85 degrees C. Several HSPs, mainly HSP 70, could be detected in the cytoplasm of these spores. An analogous HSP, the main HSP induced by increased temperature during growth, belongs to the GroEL group according to its N-terminal sequence. The identity of this protein was confirmed by Western blot (immunoblot) analysis with polyclonal antibodies against B. subtilis GroEL. Sporangia treated by heat shock immediately or 240 min after exponential phase also synthesized HSPs, but none of them could be detected in the spores in an appreciable amount. These spores showed only a slightly increased heat resistance.

• MeSH
• Autoradiography MeSH
• Bacillus megaterium growth & development   metabolism   physiology   MeSH
• Bacterial Proteins biosynthesis   isolation & purification   MeSH
• Time Factors MeSH
• Chaperonin 60 MeSH
• Cytoplasm metabolism   MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Kinetics MeSH
• Molecular Sequence Data MeSH
• Molecular Weight MeSH
• Heat-Shock Proteins biosynthesis   isolation & purification   MeSH
• Sulfur Radioisotopes MeSH
• Amino Acid Sequence MeSH
• Sulfates metabolism   MeSH
• Spores, Bacterial physiology   MeSH
• Hot Temperature MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Chaperonin 60 MeSH
• Heat-Shock Proteins MeSH
• Sulfur Radioisotopes MeSH
• Sulfates MeSH
• sodium sulfate MeSH Browser

The aim of the present study was to characterize bacteria of the genus Streptococcus isolated from the oral cavity of the guinea pig as well as to assess the significance of these microorganisms as potential veterinary and human pathogens. Sixty-two streptococcal isolates recovered from 27 clinically healthy guinea pigs were examined genotypically by sequencing the 16S rRNA and groEL genes. Among these isolates, only 13 could be assigned to a species described previously (mainly Streptococcus parasanguinis, S. mitis and S. suis), and the majority of the remaining ones differed considerably from the streptococcal species known to date (16S rRNA and groEL sequence similarities were < 97% and < 87%, respectively). Based on 16S rRNA sequences, these unidentified isolates were divided into seven groups (clades), of which clades I through III comprised most of the isolates examined and had also the widest distribution among guinea pig colonies. Upon groEL gene sequence analysis, however, members of the three clades grouped together without forming such distinct clusters. The remaining clades distinguished by 16S rRNA sequencing could also be discerned by the second gene, and they contained only a few isolates often restricted to one or a few animal colonies. The present work reveals that the guinea pig mouth is inhabited by a vast number of phylogenetically diverse, so far unrecognized populations of streptococci, most of them being apparently host-specific genomospecies. On the contrary, S. parasanguinis and S. mitis are also common human commensals and S. suis is a well-recognized zoonotic pathogen.

• MeSH
• Phylogeny MeSH
• Genetic Variation * MeSH
• Guinea Pigs MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Sequence Analysis, DNA MeSH
• Sequence Analysis MeSH
• Streptococcus * genetics   MeSH
• Animals MeSH
• Check Tag
• Guinea Pigs MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• RNA, Ribosomal, 16S MeSH

AIMS: To find out membrane tolerance strategy to ethanol in Bacillus subtilis that possesses a powerful system of protection against environmental stresses. METHODS AND RESULTS: Cytoplasmic membranes of B. subtilis were severely affected by even short-term exposure to 3% (v/v) ethanol: the growth rate and membrane protein synthesis were markedly reduced, and no adaptive alterations in phospholipids were detected. Simultaneously, steady-state DPH fluorescence anisotropy (r(ss)) showed that the membrane rigidity increased substantially. Analysis of the membrane phosphoproteome using in vitro labelling with [γ-(32) P]ATP revealed the association of DnaK and GroEL chaperones with membrane, indicating a stress induction process. Upon a long-term 3% (v/v) ethanol stress, the cell growth accelerated slightly and the composition of polar head groups and fatty acids of membrane phospholipids underwent an extensive reconstruction. Correspondingly, membrane fluidity turned back to the original r(ss) values of the control cells. CONCLUSIONS: In B. subtilis, the adaptive response to short-term ethanol stress comprises the recruitment of molecular chaperones on the impaired membrane structure; consequently, the phospholipid synthesis is restored and membrane fluidity adapts properly to the continuing ethanol stress. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings underline the role of membrane lipids in establishing tolerance towards ethanol and also suggest the contribution of molecular chaperones to the membrane and cell recovery.

• MeSH
• Bacillus subtilis drug effects   growth & development   physiology   MeSH
• Bacterial Proteins metabolism   MeSH
• Cell Membrane chemistry   metabolism   MeSH
• Ethanol metabolism   pharmacology   MeSH
• Membrane Fluidity drug effects   MeSH
• Fluorescence Polarization MeSH
• Phospholipids metabolism   MeSH
• Stress, Physiological MeSH
• Fatty Acids analysis   MeSH
• Membrane Lipids chemistry   metabolism   MeSH
• Molecular Chaperones metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Ethanol MeSH
• Phospholipids MeSH
• Fatty Acids MeSH
• Membrane Lipids MeSH
• Molecular Chaperones MeSH

Fleas (95 Pulex irritans, 50 Ctenocephalides felis, 45 Ctenocephalides canis) and ixodid ticks (223 ixodes ricinus, 231 Dermacentor reticulatus, 204 Haemaphysalis concinna) were collected in Hungary and tested, in assays based on PCR, for Bartonella infection. Low percentages of P. irritans (4.2%) and C. felis (4.0%) were found to be infected. The groEL sequences of the four isolates from P. irritans were different from all the homologous sequences for bartonellae previously stored in GenBank but closest to those of Bartonella sp. SE-Bart-B (sharing 96% identities). The groEL sequences of the two isolates from C. felis were identical with those of the causative agents of cat scratch disease, Bartonella henselae and Bartonella clarridgeiae, respectively. The pap31 sequences of B. henselae amplified from Hungarian fleas were identical with that of Marseille strain. No Bartonella-specific amplification products were detected in C. canis, I. ricinus, D. reticulatus and H. concinna pools.

• MeSH
• Bartonella genetics   growth & development   MeSH
• Chaperonin 60 chemistry   genetics   MeSH
• Arthropod Vectors microbiology   MeSH
• DNA, Bacterial chemistry   genetics   MeSH
• Insect Vectors microbiology   MeSH
• Bartonella Infections microbiology   MeSH
• Ticks microbiology   MeSH
• Cats MeSH
• Foxes MeSH
• Molecular Sequence Data MeSH
• Polymerase Chain Reaction MeSH
• Base Sequence MeSH
• Sequence Analysis, DNA MeSH
• Siphonaptera microbiology   MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Hungary MeSH
• Names of Substances
• Chaperonin 60 MeSH
• DNA, Bacterial MeSH

BACKGROUND: Anaplasma phagocytophilum is currently regarded as a single species. However, molecular studies indicate that it can be subdivided into ecotypes, each with distinct but overlapping transmission cycle. Here, we evaluate the interactions between and within clusters of haplotypes of the bacterium isolated from vertebrates and ticks, using phylogenetic and network-based methods. METHODS: The presence of A. phagocytophilum DNA was determined in ticks and vertebrate tissue samples. A fragment of the groEl gene was amplified and sequenced from qPCR-positive lysates. Additional groEl sequences from ticks and vertebrate reservoirs were obtained from GenBank and through literature searches, resulting in a dataset consisting of 1623 A. phagocytophilum field isolates. Phylogenetic analyses were used to infer clusters of haplotypes and to assess phylogenetic clustering of A. phagocytophilum in vertebrates or ticks. Network-based methods were used to resolve host-vector interactions and their relative importance in the segregating communities of haplotypes. RESULTS: Phylogenetic analyses resulted in 199 haplotypes within eight network-derived clusters, which were allocated to four ecotypes. The interactions of haplotypes between ticks, vertebrates and geographical origin, were visualized and quantified from networks. A high number of haplotypes were recorded in the tick Ixodes ricinus. Communities of A. phagocytophilum recorded from Korea, Japan, Far Eastern Russia, as well as those associated with rodents had no links with the larger set of isolates associated with I. ricinus, suggesting different evolutionary pressures. Rodents appeared to have a range of haplotypes associated with either Ixodes trianguliceps or Ixodes persulcatus and Ixodes pavlovskyi. Haplotypes found in rodents in Russia had low similarities with those recorded in rodents in other regions and shaped separate communities. CONCLUSIONS: The groEl gene fragment of A. phagocytophilum provides information about spatial segregation and associations of haplotypes to particular vector-host interactions. Further research is needed to understand the circulation of this bacterium in the gap between Europe and Asia before the overview of the speciation features of this bacterium is complete. Environmental traits may also play a role in the evolution of A. phagocytophilum in ecotypes through yet unknown relationships.

• Keywords
• Anaplasma phagocytophilum, Ixodidae, Molecular epidemiology, Network analysis, Ticks, Transmission dynamics,
• MeSH
• Anaplasma phagocytophilum genetics   isolation & purification   MeSH
• Chaperonin 60 genetics   MeSH
• Ecotype MeSH
• Phylogeny * MeSH
• Haplotypes MeSH
• Ixodes microbiology   MeSH
• Evolution, Molecular * MeSH
• Vertebrates microbiology   MeSH
• Biota * MeSH
• Geography MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Asia MeSH
• Europe MeSH
• Names of Substances
• Chaperonin 60 MeSH

All organisms have the capacity to sense and respond to environmental changes. These signals often involve the use of second messengers such as cyclic adenosine monophosphate (cAMP). This second messenger is widely distributed among organisms and coordinates gene expression related with pathogenesis, virulence, and environmental adaptation. Genomic analysis in Mycobacterium tuberculosis has identified 16 adenylyl cyclases (AC) and one phosphodiesterase, which produce and degrade cAMP, respectively. To date, ten AC have been biochemically characterized and only one (Rv0386) has been found to be important during murine infection with M. tuberculosis. Here, we investigated the impact of hsp60-driven Rv2212 gene expression in Mycobacterium bovis Bacillus Calmette-Guerin (BCG) during growth in vitro, and during macrophage and mice infection. We found that hsp60-driven expression of Rv2212 resulted in an increased capacity of replication in murine macrophages but an attenuated phenotype in lungs and spleen when administered intravenously in mice. Furthermore, this strain displayed an altered proteome mainly affecting proteins associated with stress conditions (bfrB, groEL-2, DnaK) that could contribute to the attenuated phenotype observed in mice.

• MeSH
• Adenylyl Cyclases genetics   metabolism   MeSH
• Bacterial Proteins genetics   metabolism   MeSH
• Cell Line MeSH
• Chaperonin 60 genetics   metabolism   MeSH
• Humans MeSH
• Macrophages microbiology   MeSH
• Mycobacterium bovis genetics   metabolism   pathogenicity   MeSH
• Mycobacterium tuberculosis enzymology   genetics   pathogenicity   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Lung microbiology   MeSH
• Proteome genetics   metabolism   MeSH
• Spleen microbiology   MeSH
• Tuberculosis microbiology   MeSH
• Virulence MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Adenylyl Cyclases MeSH
• Bacterial Proteins MeSH
• Chaperonin 60 MeSH
• Proteome MeSH

OBJECTIVE: Summarization of recent knowledge on heat shock proteins (HSPs) of human and bacterial (chlamydial) origin and their participation in fertility disturbances. DESIGN: Review article for training of physicians (gynecologists and obstetricians). SETTING: Veterinary Research Institute, Brno. METHOD AND RESULTS: The subject of the study is heat shock protein--hsp60 as a significant epitope Chlamydia trachomatis. Heat shock proteins are induced as a response to various stress insults from external environment (hyperthermy, UV radiation, free oxygen radicals, heavy metals, ethanol etc.) and certain processes related to the cell cycle. Sensitization with the heat shock protein Chlamydia trachomatis and subsequent excretion of highly homologous human heat shock protein are co-operating factors in the development of fertility disturbances. Significant levels of IgA antibodies to hsp60 occur in cervical mucus of women and in seminal plasma of men with fertility disturbances. CONCLUSION: Preceding infection C. trachomatis and resulting sensitization with chlamydial heat shock protein indicate an unfavourable prognosis of the reproductive outcome and impairs the perspective of a successful in vitro fertilization.

• MeSH
• Chaperonin 60 immunology   MeSH
• Chlamydia trachomatis immunology   MeSH
• Epitopes immunology   MeSH
• Infertility immunology   microbiology   MeSH
• Humans MeSH
• Antibodies, Bacterial analysis   MeSH
• Pregnancy MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Chaperonin 60 MeSH
• Epitopes MeSH
• Antibodies, Bacterial MeSH