Comparative study of the life cycle dependent post-translation modifications of protein synthesis elongation factor Tu present in the membrane proteome of streptomycetes and mycobacteria

Holub, Martin; Bezoušková, Silvia; Petráčková, Denisa; Kalachová, Ľubica; Kofroňová, Olga; Benada, Oldřich; Weiser, Jaroslav

doi:10.1007/s12223-010-0029-2

Cited by 4 publications

(4 citation statements)

References 38 publications

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“…We have recently shown that, in other Streptomycetes strains, EF-Tu is present in various isoforms differing in pI; these isoforms are present among both cytoplasm and membrane-associated factor (Nguyen et al 2005a;Holub et al 2010). For example, in dormant spores of S. coelicolor, which is a close relative of S. lividans, we observed two EF-Tu isoforms that were present in an equimolar ratio (Holub et al 2007); this result is similar to that observed in this study (Figs.…”

Section: Discussionsupporting

confidence: 90%

“…The samples (100 μg of proteins or 500,000 cpm of pulse-labelled proteins) were absorbed into IPG strips (Immobiline DryStrips, 180 mm) with a nonlinear pH 4-7 gradient (Amersham Biosciences, UK) and rehydrated overnight. The electrophoresis was performed as described previously (Holub et al 2010). The gels were silver-stained according to Rabilloud (1992).…”

Section: D Electrophoresismentioning

confidence: 99%

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The use of glass beads cultivation system to study the global effect of the ppk gene inactivation in Streptomyces lividans

et al. 2011

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The glass beads cultivation system developed in our laboratory for physiological studies of filamentous microorganisms supports differentiation and allows complete recovery of bacterial colonies and their natural products from cultivation plates. Here, we used this system to study the global effect of ppk gene disruption in Streptomyces lividans. The ppk encoding the enzyme polyphosphate kinase (P) catalyses the reversible polymerisation of gamma phosphate of ATP to polyphosphates. The resulting are phosphate and energy stock polymers. Because P activity impacts the overall energetic state of the cell, it is also connected to secondary metabolite (e.g. antibiotic) biosynthesis. We analysed the global effects of the disruption of this gene including its influence on the production of pigmented antibiotics, on morphological differentiation, on the levels of ATP and on the whole cytoplasmic protein expression pattern of S. lividans. We observed that the S. lividans ppk mutant produced antibiotics earlier and in greater amount than the wild-type (wt) strain. On the other hand, we did not observe any obvious effect on colony morphological development. In agreement with the function of Ppk, we detected much lower levels of ATP in ppk- mutant than in the wt strain. Proteomic analysis revealed that the genes that were influenced by ppk inactivation included enzymes involved in carbon or nitrogen metabolism, phosphate transport and components of the cell translational machinery. We showed that the synthesis of translation elongation factor Tu is during sporulation much higher in ppk- mutant than in wild-type strain.

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Section: Discussionsupporting

confidence: 90%

Section: D Electrophoresismentioning

confidence: 99%

The use of glass beads cultivation system to study the global effect of the ppk gene inactivation in Streptomyces lividans

et al. 2011

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“…In bacteria, some ribosomal proteins may have modified functions beyond the translation of mRNA into protein (79). Similarly, elongation factor Tu has been identified both as a surface-exposed protein and an essential virulence factor for several bacteria (14,28,35,52). Ribosomes may also be complexed with protein-translocating channels, making them partially exposed to the extracellular environment, which may also cause them to be detected in screens of surface-exposed proteins (57).…”

Section: Discussionmentioning

confidence: 99%

Surface Proteome of “Mycobacterium avium subsp. hominissuis” during the Early Stages of Macrophage Infection

et al. 2012

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ABSTRACT“Mycobacterium aviumsubsp.hominissuis” is a robust and pervasive environmental bacterium that can cause opportunistic infections in humans. The bacterium overcomes the host immune response and is capable of surviving and replicating within host macrophages. Little is known about the bacterial mechanisms that facilitate these processes, but it can be expected that surface-exposed proteins play an important role. In this study, the selective biotinylation of surface-exposed proteins, streptavidin affinity purification, and shotgun mass spectrometry were used to characterize the surface-exposed proteome ofM. aviumsubsp.hominissuis. This analysis detected more than 100 proteins exposed at the bacterial surface ofM. aviumsubsp.hominissuis. Comparisons of surface-exposed proteins between conditions simulating early infection identified several groups of proteins whose presence on the bacterial surface was either constitutive or appeared to be unique to specific culture conditions. This proteomic profile facilitates an improved understanding ofM. aviumsubsp.hominissuisand how it establishes infection. Additionally, surface-exposed proteins are excellent targets for the host adaptive immune system, and their identification can inform the development of novel treatments, diagnostic tools, and vaccines for mycobacterial disease.

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“…The beads are immersed in a liquid medium that allows for easy modification or replacement of nutrients and growth factors, as well as radioactive labelling of proteins and other macromolecules. Cultivation on beads enables studies of differentiation in streptomycetes using molecular biology and global 'omics' techniques, along with the analysis of produced secondary metabolites such as undecylprodigiosin in Streptomyces lividans (Holub et al, 2010;Nezbedova et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Surface hydrophobicity and roughness influences the morphology and biochemistry of streptomycetes during attached growth and differentiation

Petráčková

Buriánková

Tesařová

et al. 2013

FEMS Microbiol Lett

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Streptomycetes, soil-dwelling mycelial bacteria, can colonise surface of organic soil debris and soil particles. We analysed the effects of two different inert surfaces, glass and zirconia/silica, on the growth and antibiotic production in Streptomyces granaticolor. The surfaces used were in the form of microbeads and were surrounded by liquid growth media. Following the production of the antibiotic granaticin, more biomass was formed as well as a greater amount of antibiotic per milligram of protein on the glass beads than on the zirconia/silica beads. Comparison of young mycelium (6 h) proteomes, obtained from the cultures attached to the glass and zirconia/silica beads, revealed three proteins with altered expression levels (dihydrolipoamide dehydrogenase, amidophosphoribosyltransferase and cystathionine beta-synthase) and one unique protein (glyceraldehyde-3-phosphate dehydrogenase) that was present only in cells grown on glass beads. All of the identified proteins function primarily as cytoplasmic enzymes involved in different parts of metabolism; however, in several microorganisms, they are exposed on the cell surface and have been shown to be involved in adhesion or biofilm formation.

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Comparative study of the life cycle dependent post-translation modifications of protein synthesis elongation factor Tu present in the membrane proteome of streptomycetes and mycobacteria

Cited by 4 publications

References 38 publications

The use of glass beads cultivation system to study the global effect of the ppk gene inactivation in Streptomyces lividans

The use of glass beads cultivation system to study the global effect of the ppk gene inactivation in Streptomyces lividans

Surface Proteome of “Mycobacterium avium subsp. hominissuis” during the Early Stages of Macrophage Infection

Surface hydrophobicity and roughness influences the morphology and biochemistry of streptomycetes during attached growth and differentiation

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