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.

• MeSH
• adenosintrifosfát biosyntéza   MeSH
• antibakteriální látky biosyntéza   MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• fosfotransferasy s fosfátovou skupinou jako akceptorem genetika   metabolismus   MeSH
• kultivační techniky přístrojové vybavení   metody   MeSH
• molekulární sekvence - údaje MeSH
• regulace genové exprese u bakterií MeSH
• Streptomyces lividans enzymologie   genetika   růst a vývoj   metabolismus   MeSH
• umlčování genů * MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenosintrifosfát MeSH
• antibakteriální látky MeSH
• bakteriální proteiny MeSH
• fosfotransferasy s fosfátovou skupinou jako akceptorem MeSH
• polyphosphate kinase MeSH Prohlížeč

We present the results of analysis of membrane phosphoproteomes from individual morphological stages of Streptomyces coelicolor that reflect developmentally dependent heterogeneity and phosphorylation of intrinsic and externally added purified Strepomyces aureofaciens EF-Tu. Fast growing nonpathogenic Mycobacterium smegmatis was used as a non-differentiating actinomycetes comparative model. Streptomycetes membrane fraction was found to contain protein kinase(s) catalyzing phosphorylation of both its own and an externally added EF-Tu, whereas Mycobacterium membrane fraction contains protein kinase phosphorylating only its own EF-Tu.

• MeSH
• buněčná membrána chemie   enzymologie   metabolismus   MeSH
• elongační faktor Tu izolace a purifikace   metabolismus   MeSH
• fosforylace MeSH
• Mycobacterium smegmatis chemie   enzymologie   metabolismus   MeSH
• posttranslační úpravy proteinů * MeSH
• proteinkinasy izolace a purifikace   metabolismus   MeSH
• Streptomyces chemie   enzymologie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• elongační faktor Tu MeSH
• proteinkinasy MeSH

In vitro phosphorylation of EF-Tu was shown in cell-free extract from dormant spores of Streptomyces coelicolor by a protein kinase present in spores. EF-Tu phosphorylation was observed on both intrinsic S. coelicolor factor and externally added purified EF-Tu from S. aureofaciens, on two isoforms. Putative serine and threonine residues as potential phosphorylation targets were determined in primary sequence and demonstrated on 3D structure model of EF-Tu.

• MeSH
• 2D gelová elektroforéza MeSH
• elongační faktor Tu chemie   izolace a purifikace   metabolismus   MeSH
• fosforylace MeSH
• molekulární modely MeSH
• molekulární sekvence - údaje MeSH
• proteinkinasy metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční seřazení MeSH
• spory bakteriální enzymologie   metabolismus   MeSH
• Streptomyces aureofaciens metabolismus   MeSH
• Streptomyces coelicolor metabolismus   MeSH
• terciární struktura proteinů MeSH
• zobrazování trojrozměrné MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• elongační faktor Tu MeSH
• proteinkinasy MeSH