Cytological and biochemical evidence for an early cell dismantling event in surface cultures of Streptomyces antibioticus

Manteca, Ángel; Fernández, Marcos; Sánchez, Jesús

doi:10.1016/j.resmic.2005.07.003

Cited by 53 publications

(65 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although most industrial processes for secondary metabolite production are performed in liquid cultures, Streptomyces strains generally do not sporulate under these conditions (6,18,22), and most authors assumed that differentiation did not take place. Recently, a detailed analysis of Streptomyces differentiation in surface and submerged cultures has been performed, describing novel aspects of the differentiation processes of this bacterium (10)(11)(12)(13)(14)(15)(16)(17). A previously unidentified compartmentalized mycelium (MI) initiates the developmental cycle and then dies following a highly ordered sequence (PCD) (10,11,14).…”

mentioning

confidence: 99%

“…The second multinucleated mycelium corresponds to the antibiotic-producing structure under surface and submerged conditions (16). The knowledge of the existence of a multinucleated mycelium (MII) which differentiates from a compartmentalized mycelium (MI) after PCD opens a whole new scenario in which to study differentiation and is crucial for the analysis of differentiation in industrial fermentations (10)(11)(12)(13)(14)(15)(16)(17).…”

mentioning

confidence: 99%

See 1 more Smart Citation

New Method for Monitoring Programmed Cell Death and Differentiation in Submerged Streptomyces Cultures

Yagüe

Manteca

Simon

et al. 2010

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

Vital stains were used in combination with fluorimetry for the elaboration of a new method to quantify Streptomyces programmed cell death, one of the key events in Streptomyces differentiation. The experimental approach described opens the possibility of designing online protocols for automatic monitoring of industrial fermentations.Streptomyces is an extremely important bacterium for industry, since approximately two-thirds of all antibiotics are synthesized by members of this genus (4). Furthermore, streptomycetes produce large numbers of eukaryotic cell differentiation inducers and apoptosis inhibitors and inducers (19,24,25). Moreover, some authors consider that bacteria with complex life cycles (streptomycetes, cyanobacteria, etc.) are the evolutionary origin of some of the protein domains involved in programmed cell death (PCD) processes, including eukaryotic apoptosis: APATPases (apoptotic ATPases), kinases, caspases, nucleases, etc. As such, these bacteria would constitute a simple and convenient model by which to study this important phenomenon (1,3,9,12,21,26).The classical Streptomyces developmental model in confluent solid cultures assumed that differentiation processes took place along the transverse axis of the cultures (bottom up): completely viable vegetative mycelia (substrate) grew on the surface and inside agar until they underwent a PCD process, after which they differentiated into a reproductive (aerial) mycelium that grew into the air (reviewed in reference 8). Although most industrial processes for secondary metabolite production are performed in liquid cultures, Streptomyces strains generally do not sporulate under these conditions (6,18,22), and most authors assumed that differentiation did not take place. Recently, a detailed analysis of Streptomyces differentiation in surface and submerged cultures has been performed, describing novel aspects of the differentiation processes of this bacterium (10-17). A previously unidentified compartmentalized mycelium (MI) initiates the developmental cycle and then dies following a highly ordered sequence (PCD) (10,11,14). Subsequently, the remaining viable segments enlarge, yielding a multinucleated mycelium (MII) that grows in successive waves that determine the characteristic complex growth curves of this microorganism. In surface cultures, two types of second mycelium were defined, based on the absence (in early development) or presence (in late development) of the hydrophobic layers characteristic of aerial hyphae (5). The traditionally denominated substrate (vegetative) mycelium corresponds, in fact, to the early second multinucleated mycelium that still lacks the hydrophobic layers coating the aerial mycelium (15). We proposed that the first compartmentalized mycelium fulfills the true vegetative role in Streptomyces development in soil (17). According to this scheme, the second early and late multinucleated mycelia should be considered jointly as part of the reproductive phase, since they are destined to sporulate (17). The second multinuclea...

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

New Method for Monitoring Programmed Cell Death and Differentiation in Submerged Streptomyces Cultures

Yagüe

Manteca

Simon

et al. 2010

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Culture samples were obtained and processed for microscopy at different incubation time points, as previously described (Manteca et al 2006). Cells were stained with propidium iodide and SYTO 9 (LIVE/DEAD BacLight Bacterial Viability Kit, Invitrogen, L-13152).…”

Section: Viability Stainingmentioning

confidence: 99%

“…Cells were stained with propidium iodide and SYTO 9 (LIVE/DEAD BacLight Bacterial Viability Kit, Invitrogen, L-13152). The samples were observed under a Leica TCS-SP2-AOBS confocal laser scanning microscope at wavelengths of 488 and 568 nm excitation and 530 nm (green) or 640 nm (red) emissions (Manteca et al 2006). …”

Section: Viability Stainingmentioning

confidence: 99%

New ΦBT1 site-specific integrative vectors with neutral phenotype in Streptomyces

Gonzalez-Quiñonez

López-García

Yagüe

et al. 2016

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Integrative plasmids are one of the best options to introduce genes in low copy and in a stable form into bacteria. The ΦC31-derived plasmids constitute the most common integrative vectors used in Streptomyces. They integrate at different positions (attB and pseudo-attB sites) generating different mutations. The less common ΦBT1-derived vectors integrate at the unique attB site localized in the SCO4848 gene (S. coelicolor genome) or their orthologues in other streptomycetes. This work demonstrates that disruption of SCO4848 generates a delay in spore germination. SCO4848 is cotranscribed with SCO4849, and the spore germination phenotype is complemented by SCO4849. Plasmids pNG1-4 were created by modifying the ΦBT1 integrative vector pMS82 by introducing a copy of SCO4849 under the control of the promoter region of SCO4848. pNG2 and pNG4 also included a copy of the P ermE * in order to facilitate gene overexpression. pNG3 and pNG4 harboured a copy of the bla gene (ampicillin resistance) to facilitate selection in E. coli. pNG1-4 are the only integrative vectors designed to produce a neutral phenotype when they are integrated into the Streptomyces genome. The experimental approach developed in this work can be applied to create phenotypically neutral integrative plasmids in other bacteria.

show abstract

“…This round of PCD is thought to correspond to the short period of growth cessation called the transition phase that takes places during the exponential growth and is followed by a slower regain of growth before entry into stationary phase (Granozzi et al 1990;Puglia et al 1995;Manteca et al 2005). The cell death paroxysm at the transition phase causes intensive release of intracellular compounds into the extracellular environment (Manteca et al 2006). The latter are thought to serve as nutritive sources for surviving vegetative filaments to grow and differentiate in the aerial structures (Mendez et al 1985).…”

Section: Introductionmentioning

confidence: 99%

Extracellular sugar phosphates are assimilated by Streptomyces in a PhoP-dependent manner

Tenconi

Jourdan

Motté

et al. 2012

Antonie van Leeuwenhoek

View full text Add to dashboard Cite

Filamentous microorganisms of the bacterial genus Streptomyces have a complex life cycle that includes physiological and morphological differentiations. It is now fairly well accepted that lysis of Streptomyces vegetative mycelium induced by programmed cell death (PCD) provides the required nutritive sources for the bacterium to erect sporeforming aerial hyphae. However, little is known regarding cellular compounds released during PCD and the contribution of these molecules to the feeding of surviving cells in order to allow them to reach the late stages of the developmental program. In this work we assessed the effect of extracellular sugar phosphates (that are likely to be released in the environment upon cell lysis) on the differentiation processes.We demonstrated that the supply of phosphorylated sugars, under inorganic phosphate limitation, delays the occurrence of the second round of PCD, blocks streptomycetes life cycle at the vegetative state and inhibits antibiotic production. The mechanism by which sugar phosphates affect development was shown to involve genes of the Pho regulon that are under the positive control of the two component system PhoR/PhoP. Indeed, the inactivation of the response regulator phoP of Streptomyces lividans prevented the 'sugar phosphate effect' whereas the S. lividans ppk (polyphosphate kinase) deletion mutant, known to overexpress the Pho regulon, presented an enhanced response to phosphorylated sugars.

show abstract

Cytological and biochemical evidence for an early cell dismantling event in surface cultures of Streptomyces antibioticus

Cited by 53 publications

References 39 publications

New Method for Monitoring Programmed Cell Death and Differentiation in Submerged Streptomyces Cultures

New Method for Monitoring Programmed Cell Death and Differentiation in Submerged Streptomyces Cultures

New ΦBT1 site-specific integrative vectors with neutral phenotype in Streptomyces

Extracellular sugar phosphates are assimilated by Streptomyces in a PhoP-dependent manner

Contact Info

Product

Resources

About