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1.
Proc Natl Acad Sci U S A ; 117(25): 14444-14452, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32513721

RESUMO

Chemical-induced spores of the Gram-negative bacterium Myxococcus xanthus are peptidoglycan (PG)-deficient. It is unclear how these spherical spores germinate into rod-shaped, walled cells without preexisting PG templates. We found that germinating spores first synthesize PG randomly on spherical surfaces. MglB, a GTPase-activating protein, forms a cluster that responds to the status of PG growth and stabilizes at one future cell pole. Following MglB, the Ras family GTPase MglA localizes to the second pole. MglA directs molecular motors to transport the bacterial actin homolog MreB and the Rod PG synthesis complexes away from poles. The Rod system establishes rod shape de novo by elongating PG at nonpolar regions. Thus, similar to eukaryotic cells, the interactions between GTPase, cytoskeletons, and molecular motors initiate spontaneous polarization in bacteria.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas Ativadoras de GTPase/metabolismo , Myxococcus xanthus/citologia , Peptidoglicano/metabolismo , Esporos Bacterianos/crescimento & desenvolvimento , Polaridade Celular , Parede Celular/metabolismo , Parede Celular/ultraestrutura , Microscopia Eletrônica , Morfogênese , Myxococcus xanthus/crescimento & desenvolvimento , Myxococcus xanthus/metabolismo , Myxococcus xanthus/ultraestrutura , Peptidoglicano/genética , Esporos Bacterianos/metabolismo , Esporos Bacterianos/ultraestrutura
2.
Cells ; 8(6)2019 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-31163575

RESUMO

Two unrecognizable strains of the same bacterial species form a distinct colony boundary. During growth as colonies, Myxococcus xanthus uses multiple factors to establish cooperation between recognized strains and prevent interactions with unrecognized strains of the same species. Here, ΔMXAN_0049 is a mutant strain deficient in immunity for the paired nuclease gene, MXAN_0050, that has a function in the colony-merger incompatibility of Myxococcus xanthus DK1622. With the aim to investigate the factors involved in boundary formation, a proteome and metabolome study was employed. Visualization of the boundary between DK1622 and ΔMXAN_0049 was done scanning electron microscope (SEM), which displayed the presence of many damaged cells in the boundary. Proteome analysis of the DK1622- boundary disclosed many possible proteins, such as cold shock proteins, cell shape-determining protein MreC, along with a few pathways, such as RNA degradation, phenylalanine, tyrosine and tryptophan biosynthesis, and Type VI secretion system (T6SS), which may play major roles in the boundary formation. Metabolomics studies revealed various secondary metabolites that were significantly produced during boundary formation. Overall, the results concluded that multiple factors participated in the boundary formation in M. xanthus, leading to cellular damage that is helpful in solving the mystery of the boundary formation mechanism.


Assuntos
Metabolômica/métodos , Myxococcus xanthus/crescimento & desenvolvimento , Myxococcus xanthus/metabolismo , Proteômica/métodos , Sistemas de Secreção Bacterianos , Contagem de Colônia Microbiana , Regulação para Baixo , Viabilidade Microbiana , Myxococcus xanthus/ultraestrutura , Mapas de Interação de Proteínas , Proteoma/metabolismo , Metabolismo Secundário , Regulação para Cima
3.
EMBO Rep ; 18(7): 1090-1099, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28487352

RESUMO

The type VI secretion system (T6SS) is a versatile molecular weapon used by many bacteria against eukaryotic hosts or prokaryotic competitors. It consists of a cytoplasmic bacteriophage tail-like structure anchored in the bacterial cell envelope via a cytoplasmic baseplate and a periplasmic membrane complex. Rapid contraction of the sheath in the bacteriophage tail-like structure propels an inner tube/spike complex through the target cell envelope to deliver effectors. While structures of purified contracted sheath and purified membrane complex have been solved, because sheaths contract upon cell lysis and purification, no structure is available for the extended sheath. Structural information about the baseplate is also lacking. Here, we use electron cryotomography to directly visualize intact T6SS structures inside Myxococcus xanthus cells. Using sub-tomogram averaging, we resolve the structure of the extended sheath and membrane-associated components including the baseplate. Moreover, we identify novel extracellular bacteriophage tail fiber-like antennae. These results provide new structural insights into how the extended sheath prevents premature disassembly and how this sophisticated machine may recognize targets.


Assuntos
Myxococcus xanthus/ultraestrutura , Sistemas de Secreção Tipo VI/ultraestrutura , Bacteriófagos/ultraestrutura , Microscopia Crioeletrônica/instrumentação , Microscopia Crioeletrônica/métodos , Microscopia Intravital/instrumentação , Microscopia Intravital/métodos , Estrutura Molecular , Myxococcus xanthus/química , Myxococcus xanthus/citologia , Ligação Proteica , Multimerização Proteica , Sistemas de Secreção Tipo VI/química
4.
Nat Microbiol ; 2: 16269, 2017 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-28165453

RESUMO

Type IV pili (T4P) are filamentous appendages found on many Bacteria and Archaea. They are helical fibres of pilin proteins assembled by a multi-component macromolecular machine we call the basal body. Based on pilin features, T4P are classified into type IVa pili (T4aP) and type IVb pili (T4bP)1,2. T4aP are more widespread and are involved in cell motility3, DNA transfer4, host predation5 and electron transfer6. T4bP are less prevalent and are mainly found in enteropathogenic bacteria, where they play key roles in host colonization7. Following similar work on T4aP machines8,9, here we use electron cryotomography10 to reveal the three-dimensional in situ structure of a T4bP machine in its piliated and non-piliated states. The specific machine we analyse is the Vibrio cholerae toxin-coregulated pilus machine (TCPM). Although only about half of the components of the TCPM show sequence homology to components of the previously analysed Myxococcus xanthus T4aP machine (T4aPM), we find that their structures are nevertheless remarkably similar. Based on homologies with components of the M. xanthus T4aPM and additional reconstructions of TCPM mutants in which the non-homologous proteins are individually deleted, we propose locations for all eight TCPM components within the complex. Non-homologous proteins in the T4aPM and TCPM are found to form similar structures, suggesting new hypotheses for their functions and evolutionary histories.


Assuntos
Proteínas de Fímbrias/química , Fímbrias Bacterianas/química , Fímbrias Bacterianas/ultraestrutura , Vibrio cholerae/ultraestrutura , Aderência Bacteriana , Proteínas de Bactérias/análise , Proteínas de Bactérias/química , Toxina da Cólera/metabolismo , Tomografia com Microscopia Eletrônica/métodos , Proteínas de Fímbrias/análise , Fímbrias Bacterianas/genética , Modelos Moleculares , Mutação , Myxococcus xanthus/química , Myxococcus xanthus/ultraestrutura , Vibrio cholerae/química
6.
Science ; 351(6278): aad2001, 2016 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-26965631

RESUMO

Type IVa pili are filamentous cell surface structures observed in many bacteria. They pull cells forward by extending, adhering to surfaces, and then retracting. We used cryo-electron tomography of intact Myxococcus xanthus cells to visualize type IVa pili and the protein machine that assembles and retracts them (the type IVa pilus machine, or T4PM) in situ, in both the piliated and nonpiliated states, at a resolution of 3 to 4 nanometers. We found that T4PM comprises an outer membrane pore, four interconnected ring structures in the periplasm and cytoplasm, a cytoplasmic disc and dome, and a periplasmic stem. By systematically imaging mutants lacking defined T4PM proteins or with individual proteins fused to tags, we mapped the locations of all 10 T4PM core components and the minor pilins, thereby providing insights into pilus assembly, structure, and function.


Assuntos
Fímbrias Bacterianas/ultraestrutura , Myxococcus xanthus/ultraestrutura , Aderência Bacteriana , Microscopia Crioeletrônica , Fímbrias Bacterianas/genética , Microscopia Eletrônica de Transmissão , Modelos Moleculares , Mutação , Myxococcus xanthus/genética , Myxococcus xanthus/fisiologia
7.
Proc Natl Acad Sci U S A ; 112(22): E2939-46, 2015 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-26038568

RESUMO

Bacterial cells in their native environments must cope with factors that compromise the integrity of the cell. The mechanisms of coping with damage in a social or multicellular context are poorly understood. Here we investigated how a model social bacterium, Myxococcus xanthus, approaches this problem. We focused on the social behavior of outer membrane exchange (OME), in which cells transiently fuse and exchange their outer membrane (OM) contents. This behavior requires TraA, a homophilic cell surface receptor that identifies kin based on similarities in a polymorphic region, and the TraB cohort protein. As observed by electron microscopy, TraAB overexpression catalyzed a prefusion OM junction between cells. We then showed that damage sustained by the OM of one population was repaired by OME with a healthy population. Specifically, LPS mutants that were defective in motility and sporulation were rescued by OME with healthy donors. In addition, a mutant with a conditional lethal mutation in lpxC, an essential gene required for lipid A biosynthesis, was rescued by Tra-dependent interactions with a healthy population. Furthermore, lpxC cells with damaged OMs, which were more susceptible to antibiotics, had resistance conferred to them by OME with healthy donors. We also show that OME has beneficial fitness consequences to all cells. Here, in merged populations of damaged and healthy cells, OME catalyzed a dilution of OM damage, increasing developmental sporulation outcomes of the combined population by allowing it to reach a threshold density. We propose that OME is a mechanism that myxobacteria use to overcome cell damage and to transition to a multicellular organism.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Lipopolissacarídeos/metabolismo , Interações Microbianas/fisiologia , Myxococcus xanthus/fisiologia , Primers do DNA/genética , Aptidão Genética/fisiologia , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Mutagênese , Myxococcus xanthus/ultraestrutura , Reação em Cadeia da Polimerase
8.
EMBO J ; 33(17): 1896-911, 2014 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-25024436

RESUMO

Living cells compartmentalize materials and enzymatic reactions to increase metabolic efficiency. While eukaryotes use membrane-bound organelles, bacteria and archaea rely primarily on protein-bound nanocompartments. Encapsulins constitute a class of nanocompartments widespread in bacteria and archaea whose functions have hitherto been unclear. Here, we characterize the encapsulin nanocompartment from Myxococcus xanthus, which consists of a shell protein (EncA, 32.5 kDa) and three internal proteins (EncB, 17 kDa; EncC, 13 kDa; EncD, 11 kDa). Using cryo-electron microscopy, we determined that EncA self-assembles into an icosahedral shell 32 nm in diameter (26 nm internal diameter), built from 180 subunits with the fold first observed in bacteriophage HK97 capsid. The internal proteins, of which EncB and EncC have ferritin-like domains, attach to its inner surface. Native nanocompartments have dense iron-rich cores. Functionally, they resemble ferritins, cage-like iron storage proteins, but with a massively greater capacity (~30,000 iron atoms versus ~3,000 in ferritin). Physiological data reveal that few nanocompartments are assembled during vegetative growth, but they increase fivefold upon starvation, protecting cells from oxidative stress through iron sequestration.


Assuntos
Fenômenos Fisiológicos Bacterianos , Proteínas de Bactérias/metabolismo , Ferro/metabolismo , Substâncias Macromoleculares/metabolismo , Myxococcus xanthus/fisiologia , Nanopartículas/metabolismo , Estresse Oxidativo , Microscopia Crioeletrônica , Modelos Moleculares , Myxococcus xanthus/ultraestrutura , Multimerização Proteica
9.
Nat Methods ; 11(7): 737-9, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24813625

RESUMO

Cryo-electron tomography (CET) produces three-dimensional images of cells in a near-native state at macromolecular resolution, but identifying structures of interest can be challenging. Here we describe a correlated cryo-PALM (photoactivated localization microscopy)-CET method for localizing objects within cryo-tomograms to beyond the diffraction limit of the light microscope. Using cryo-PALM-CET, we identified multiple and new conformations of the dynamic type VI secretion system in the crowded interior of Myxococcus xanthus.


Assuntos
Sistemas de Secreção Bacterianos , Microscopia Crioeletrônica/métodos , Tomografia com Microscopia Eletrônica/métodos , Myxococcus xanthus/ultraestrutura , Imageamento Tridimensional/métodos , Myxococcus xanthus/química
10.
Environ Microbiol ; 16(2): 598-610, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23848955

RESUMO

The social soil bacterium, Myxococcus xanthus, displays a variety of complex and highly coordinated behaviours, including social motility, predatory rippling and fruiting body formation. Here we show that M. xanthus cells produce a network of outer membrane extensions in the form of outer membrane vesicle chains and membrane tubes that interconnect cells. We observed peritrichous display of vesicles and vesicle chains, and increased abundance in biofilms compared with planktonic cultures. By applying a range of imaging techniques, including three-dimensional (3D) focused ion beam scanning electron microscopy, we determined these structures to range between 30 and 60 nm in width and up to 5 µm in length. Purified vesicle chains consist of typical M. xanthus lipids, fucose, mannose, N-acetylglucosamine and N-acetylgalactoseamine carbohydrates and a small set of cargo protein. The protein content includes CglB and Tgl outer membrane proteins known to be transferable between cells in a contact-dependent manner. Most significantly, the 3D organization of cells within biofilms indicates that cells are connected via an extensive network of membrane extensions that may connect cells at the level of the periplasmic space. Such a network would allow the transfer of membrane proteins and other molecules between cells, and therefore could provide a mechanism for the coordination of social activities.


Assuntos
Proteínas da Membrana Bacteriana Externa/química , Biofilmes , Matriz Extracelular/ultraestrutura , Myxococcus xanthus/química , Membrana Celular/ultraestrutura , Microscopia Eletrônica de Varredura , Myxococcus xanthus/fisiologia , Myxococcus xanthus/ultraestrutura
11.
PLoS One ; 6(11): e27475, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22132103

RESUMO

Myxococcus xanthus DK1622 contains inner (IM) and outer membranes (OM) separated by a peptidoglycan layer. Integral membrane, ß-barrel proteins are found exclusively in the OM where they form pores allowing the passage of nutrients, waste products and signals. One porin, Oar, is required for intercellular communication of the C-signal. An oar mutant produces CsgA but is unable to ripple or stimulate csgA mutants to develop suggesting that it is the channel for C-signaling. Six prediction programs were evaluated for their ability to identify ß-barrel proteins. No program was reliable unless the predicted proteins were first parsed using Signal P, Lipo P and TMHMM, after which TMBETA-SVM and TMBETADISC-RBF identified ß-barrel proteins most accurately. 228 ß-barrel proteins were predicted from among 7331 protein coding regions, representing 3.1% of total genes. Sucrose density gradients were used to separate vegetative cell IM and OM fractions, and LC-MS/MS of OM proteins identified 54 ß-barrel proteins. Another class of membrane proteins, the lipoproteins, are anchored in the membrane via a lipid moiety at the N-terminus. 44 OM proteins identified by LC-MS/MS were predicted lipoproteins. Lipoproteins are distributed between the IM, OM and ECM according to an N-terminal sorting sequence that varies among species. Sequence analysis revealed conservation of alanine at the +7 position of mature ECM lipoproteins, lysine at the +2 position of IM lipoproteins, and no noticable conservation within the OM lipoproteins. Site directed mutagenesis and immuno transmission electron microscopy showed that alanine at the +7 position is essential for sorting of the lipoprotein FibA into the ECM. FibA appears at normal levels in the ECM even when a +2 lysine is added to the signal sequence. These results suggest that ECM proteins have a unique method of secretion. It is now possible to target lipoproteins to specific IM, OM and ECM locations by manipulating the amino acid sequence near the +1 cysteine processing site.


Assuntos
Proteínas de Bactérias/metabolismo , Lipoproteínas/metabolismo , Myxococcus xanthus/metabolismo , Porinas/metabolismo , Sequência de Aminoácidos , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Proteínas de Bactérias/química , Cromatografia Líquida , Biologia Computacional , Bases de Dados de Proteínas , Proteínas da Matriz Extracelular/metabolismo , Lipoproteínas/química , Espectrometria de Massas , Dados de Sequência Molecular , Myxococcus xanthus/ultraestrutura , Porinas/química , Sinais Direcionadores de Proteínas , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Transporte Proteico , Proteoma/metabolismo , Transdução de Sinais
12.
J Bacteriol ; 193(12): 3042-8, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21515768

RESUMO

Bacterial sporulation in Gram-positive bacteria results in small acid-soluble proteins called SASPs that bind to DNA and prevent the damaging effects of UV radiation. Orthologs of Bacillus subtilis genes encoding SASPs can be found in many sporulating and nonsporulating bacteria, but they are noticeably absent from spore-forming, Gram-negative Myxococcus xanthus. This is despite the fact that M. xanthus can form UV-resistant spores. Here we report evidence that M. xanthus produces its own unique group of low-molecular-weight, acid-soluble proteins that facilitate UV resistance in spores. These M. xanthus-specific SASPs vary depending upon whether spore formation is induced by starvation inside cell aggregations of fruiting bodies or is induced artificially by glycerol induction. Molecular predictions indicate that M. xanthus SASPs may have some association with the cell walls of M. xanthus spores, which may signify a different mechanism of UV protection than that seen in Gram-positive spores.


Assuntos
Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Myxococcus xanthus/fisiologia , Esporos Bacterianos/efeitos da radiação , Raios Ultravioleta , Ácidos , Eletroforese em Gel de Poliacrilamida , Mutação , Myxococcus xanthus/ultraestrutura , Esporos Bacterianos/ultraestrutura , Fatores de Tempo
13.
Mol Microbiol ; 80(4): 1031-51, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21414039

RESUMO

Bactofilins are fibre-forming bacterial cytoskeletal proteins. Here, we report the structural and biochemical characterization of MXAN_7475 (BacM), one of the four bactofilins of Myxococcus xanthus. Absence of BacM leads to a characteristic 'crooked' cell morphology and an increased sensitivity to antibiotics targeting cell wall biosynthesis. The absence of the other three bactofilins MXAN_4637-4635 (BacN-P) has no obvious phenotype. In M. xanthus, BacM exists as a 150-amino-acid full-length version and as a version cleaved before Ser28. In the cell, native BacM forms 3 nm wide fibres, which assemble into bundles forming helix-like cytoplasmic cables throughout the cell, and in a subset of cells additionally a polarly arranged lateral rod-like structure. Isolated fibres consist almost completely of the N-terminally truncated version, suggesting that the proteolytic cleavage occurs before or during fibre formation. Fusion of BacM to mCherry perturbs BacM function and cellular fibre arrangement, resulting for example in the formation of one prominent polar corkscrew-like structure per cell. Immunofluorescence staining of BacM and MreB shows that their cellular distributions are not matching. Taken together, these data suggest that rod-shaped bacteria like M. xanthus use bactofilin fibres to achieve and maintain their characteristic cell morphology and cell wall stability.


Assuntos
Proteínas de Bactérias/metabolismo , Fenômenos Fisiológicos Celulares , Proteínas do Citoesqueleto/metabolismo , Myxococcus xanthus/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/ultraestrutura , Parede Celular/metabolismo , Proteínas do Citoesqueleto/química , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/ultraestrutura , Farmacorresistência Bacteriana , Imunofluorescência , Immunoblotting , Microscopia Eletrônica , Microscopia de Fluorescência , Myxococcus xanthus/química , Myxococcus xanthus/citologia , Myxococcus xanthus/genética , Myxococcus xanthus/ultraestrutura , Ligação Proteica , Isoformas de Proteínas
14.
J Proteome Res ; 9(10): 5197-208, 2010 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-20687614

RESUMO

Social behavior in the bacterium Myxococcus xanthus relies on contact-dependent activities involving cell-cell and cell-substratum interactions. To identify outer membrane proteins that have a role in these activities, we profiled the outer membrane proteome of growing and starving cells using two strategies. First, outer membrane proteins were enriched by biotinylation of intact cells using the reagent NHS (N-hydroxysuccinimide)-PEO(12) (polyethylene oxide)-biotin with subsequent membrane solubilization and affinity chromatography. Second, the proteome of outer membrane vesicles (OMV) was determined. Comparisons of detected proteins show that these methods have different detection profiles and together provide a comprehensive view of the outer membrane proteome. From 362 proteins identified, 274 (76%) were cell envelope proteins including 64 integral outer membrane proteins and 85 lipoproteins. The majority of these proteins were of unknown function. Among integral outer membrane proteins with homologues of known function, TonB-dependent transporters comprise the largest group. Our data suggest novel functions for these transporters. Among lipoproteins with homologues of known function, proteins with hydrolytic functions comprise the largest group. The luminal load of OMV was enriched for proteins with hydrolytic functions. Our data suggest that OMV have functions in predation and possibly in transfer of intercellular signaling molecules between cells.


Assuntos
Proteínas da Membrana Bacteriana Externa/análise , Myxococcus xanthus/metabolismo , Proteoma/análise , Proteômica/métodos , Aderência Bacteriana , Biotinilação , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Cromatografia de Afinidade , Microscopia Eletrônica , Myxococcus xanthus/fisiologia , Myxococcus xanthus/ultraestrutura , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
15.
Mol Microbiol ; 74(2): 497-517, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19788540

RESUMO

Cell differentiation is widespread during the development of multicellular organisms, but rarely observed in prokaryotes. One example of prokaryotic differentiation is the gram-negative bacterium Myxococcus xanthus. In response to starvation, this gliding bacterium initiates a complex developmental programme that results in the formation of spore-filled fruiting bodies. How the cells metabolically support the necessary complex cellular differentiation from rod-shaped vegetative cells into spherical spores is unknown. Here, we present evidence that intracellular lipid bodies provide the necessary metabolic fuel for the development of spores. Formed at the onset of starvation, these lipid bodies gradually disappear until they are completely used up by the time the cells have become mature spores. Moreover, it appears that lipid body formation in M. xanthus is an important initial step indicating cell fate during differentiation. Upon starvation, two subpopulations of cells occur: cells that form lipid bodies invariably develop into spores, while cells that do not form lipid bodies end up becoming peripheral rods, which are cells that lack signs of morphological differentiation and stay in a vegetative-like state. These data indicate that lipid bodies not only fuel cellular differentiation but that their formation represents the first known morphological sign indicating cell fate during differentiation.


Assuntos
Metabolismo dos Lipídeos , Myxococcus xanthus/ultraestrutura , Esporos Bacterianos/ultraestrutura , Lipídeos/isolamento & purificação , Microscopia Eletrônica , Mutação , Myxococcus xanthus/genética , Myxococcus xanthus/crescimento & desenvolvimento , Myxococcus xanthus/metabolismo , Proteoma , Esporos Bacterianos/genética , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Bacterianos/metabolismo , Estresse Fisiológico
16.
J Bacteriol ; 191(7): 2077-82, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19168614

RESUMO

Despite the fact that most bacteria grow in biofilms in natural and pathogenic ecosystems, very little is known about the ultrastructure of their component cells or about the details of their community architecture. We used high-pressure freezing and freeze-substitution to minimize the artifacts of chemical fixation, sample aggregation, and sample extraction. As a further innovation we have, for the first time in biofilm research, used electron tomography and three-dimensional (3D) visualization to better resolve the macromolecular 3D ultrastructure of a biofilm. This combination of superb specimen preparation and greatly improved resolution in the z axis has opened a window in studies of Myxococcus xanthus cell ultrastructure and biofilm community architecture. New structural information on the chromatin body, cytoplasmic organization, membrane apposition between adjacent cells, and structure and distribution of pili and vesicles in the biofilm matrix is presented.


Assuntos
Biofilmes/crescimento & desenvolvimento , Tomografia com Microscopia Eletrônica/métodos , Imageamento Tridimensional , Myxococcus xanthus/ultraestrutura , Cromossomos Bacterianos/ultraestrutura , Vesículas Citoplasmáticas/ultraestrutura , Fímbrias Bacterianas/ultraestrutura , Myxococcus xanthus/fisiologia
18.
J Bacteriol ; 189(21): 7920-1, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17704221

RESUMO

Two models have been proposed to explain the adventurous gliding motility of Myxococcus xanthus: (i) polar secretion of slime and (ii) an unknown motor that uses cell surface adhesion complexes that form periodic attachments along the cell length. Gliding movements of the leading poles of cephalexin-treated filamentous cells were observed but not equivalent movements of the lagging poles. This demonstrates that the adventurous-motility motors are not confined to the rear of the cell.


Assuntos
Movimento/fisiologia , Myxococcus xanthus/fisiologia , Adesão Celular/fisiologia , Membrana Celular/fisiologia , Myxococcus xanthus/citologia , Myxococcus xanthus/ultraestrutura
19.
Cell Mol Life Sci ; 64(21): 2733-45, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17653507

RESUMO

Bacterial motility is essential for chemotaxis, virulence and complex social interactions leading to biofilm and fruiting body formation. Although bacterial swimming in liquids with a flagellum is well understood, little is known regarding bacterial movements across solid surfaces. Gliding motility, one such mode of locomotion, has remained largely mysterious because cells move smoothly along their long axis in the absence of any visible organelle. In this review, I discuss recent evidence that focal adhesion systems mediate gliding motility in the social bacterium Myxococcus xanthus and combine this evidence with previous work to suggest a new working hypothesis inspired from knowledge in apicomplexan parasites. I then propose experimental directions to test the model and compare it to other pre-existing models. Finally, evidence on gliding mechanisms of selected organisms are presented to ask whether some features of the model have precedents in other bacteria and whether this complex biological process could be explained by a single mechanism or involves multiple distinct mechanisms.


Assuntos
Myxococcus xanthus/fisiologia , Aderência Bacteriana/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/fisiologia , Citoesqueleto/fisiologia , Adesões Focais/fisiologia , Adesões Focais/ultraestrutura , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Modelos Biológicos , Movimento/fisiologia , Myxococcus xanthus/genética , Myxococcus xanthus/ultraestrutura , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
20.
J Bacteriol ; 189(13): 4887-98, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17483223

RESUMO

Myxococcus xanthus is a soil bacterium that undergoes a unique life cycle among the prokaryotes upon starvation, which includes the formation of macroscopic structures, the fruiting bodies, and the differentiation of vegetative rods into coccoid myxospores. This peculiarity offers the opportunity to study the copper response in this bacterium in two different stages. In fact, M. xanthus vegetative rods exhibit 15-fold-greater resistance against copper than developing cells. However, cells pre-adapted to this metal reach the same levels of resistance during both stages. Analysis of the M. xanthus genome reveals that many of the genes involved in copper resistance are redundant, three of which encode proteins of the multicopper oxidase family (MCO). Each MCO gene exhibits a different expression profile in response to external copper addition. Promoters of cuoA and cuoB respond to Cu(II) ions during growth and development; however, they show a 10-fold-increased copper sensitivity during development. The promoter of cuoC shows copper-independent induction upon starvation, but it is copper up-regulated during growth. Phenotypic analyses of deletion mutants reveal that CuoB is involved in the primary copper-adaptive response; CuoA and CuoC are necessary for the maintenance of copper tolerance; and CuoC is required for normal development. These roles seem to be carried out through cuprous oxidase activity.


Assuntos
Proteínas de Bactérias/genética , Perfilação da Expressão Gênica , Myxococcus xanthus/genética , Oxirredutases/genética , Sequência de Aminoácidos , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/fisiologia , Cobre/metabolismo , Cobre/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Genoma Bacteriano , Isoenzimas/genética , Isoenzimas/metabolismo , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Mutação , Myxococcus xanthus/crescimento & desenvolvimento , Myxococcus xanthus/ultraestrutura , Oxirredutases/metabolismo , Homologia de Sequência de Aminoácidos
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