FtsZ phosphorylation pleiotropically affects Z-ladder formation, antibiotic production, and morphogenesis in Streptomyces coelicolor

Abstract

The GTPase FtsZ forms the cell division scaffold in bacteria, which mediates the recruitment of the other components of the divisome. Streptomycetes undergo two different forms of cell division. Septa without detectable peptidoglycan divide the highly compartmentalised young hyphae during early vegetative growth, and cross-walls are formed that dissect the hyphae into long multinucleoid compartments in the substrate mycelium, while ladders of septa are formed in the aerial hyphae that lead to chains of uninucleoid spores. In a previous study, we analysed the phosphoproteome of Streptomyces coelicolor and showed that FtsZ is phosphorylated at Ser 317 and Ser389. Substituting Ser–Ser for either Glu–Glu (mimicking phosphorylation) or Ala–Ala (mimicking non-phosphorylation) hinted at changes in antibiotic production. Here we analyse development, colony morphology, spore resistance, and antibiotic production in FtsZ knockout mutants expressing FtsZ alleles mimicking Ser319 and Ser387 phosphorylation and non-phosphorylation: AA (no phosphorylation), AE, EA (mixed), and EE (double phosphorylation). The FtsZ-eGFP AE, EA and EE alleles were not able to form observable FtsZ-eGFP ladders when they were expressed in the S. coelicolor wild-type strain, whereas the AA allele could form apparently normal eGFP Z-ladders. The FtsZ mutant expressing the FtsZ EE or EA or AE alleles is able to sporulate indicating that the mutant alleles are able to form functional Z-rings leading to sporulation when the wild-type FtsZ gene is absent. The four mutants were pleiotropically affected in colony morphogenesis, antibiotic production, substrate mycelium differentiation and sporulation (sporulation timing and spore resistance) which may be an indirect result of the effect in sporulation Z-ladder formation. Each mutant showed a distinctive phenotype in antibiotic production, single colony morphology, and sporulation (sporulation timing and spore resistance) indicating that the different FtsZ phosphomimetic alleles led to different phenotypes. Taken together, our data provide evidence for a pleiotropic effect of FtsZ phosphorylation in colony morphology, antibiotic production, and sporulation.

Elicitation of Cryptic Secondary Metabolites and Antibacterial Activities from Mangrove and Cave Soil Actinomycetes

“…It is presumably because these genes are seldom expressed or expressed poorly, with the majority remaining phenotypically silent under conventional screening conditions (Abdelmohsen et al, 2015;Begani et al, 2018). A single Streptomyces strain is estimated to produce around 4 compounds but encodes 20-50 secondary metabolite pathways (Belknap et al, 2020;Caboche, 2014;Yagüe et al, 2022). Conversely, other actinomycete families, including Corynebacteriales, Micromonosporaceae, Streptosporanqineae, as well as Pseudonocardiales, possess approximately 8.4, 13.3, 15.0, and 19.8 secondary metabolite biosynthetic gene clusters (BGCs) per genome and only a fraction of the compounds have been isolated using standard fermentation conditions (Doroghazi et al, 2014).…”