Physiology and Transcriptional Analysis of ppGpp-Related Regulatory Effects in Streptomyces diastatochromogenes 1628

Song, Yang; Zhang, Xiangli; Zhang, Zixuan; Shentu, Xuping; Yu, Xiaoping

doi:10.1128/spectrum.01200-22

Cited by 3 publications

(3 citation statements)

References 29 publications

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“…The primers are listed in Supplementary Table S1. S. diastatochromogenes 1628 has been deposited in the China General Microbiological Culture Collection (CGMCC 2060), and was isolated from Tianmu Mountain, Zhejiang Province, in previous work [27]. The strain was grown at 28 • C in a glucose yeast mineral (GYM) medium [28] for a morphological analysis.…”

Section: Strains Plasmids Primers and Culture Conditionsmentioning

confidence: 99%

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CdgB Regulates Morphological Differentiation and Toyocamycin Production in Streptomyces diastatochromogenes 1628

Wang,

Zhang,

et al. 2024

IJMS

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Bis (3′,5′)-cyclic diguanylic acid (c-di-GMP) is a ubiquitous second messenger that controls several metabolic pathways in bacteria. In Streptomyces, c-di-GMP is associated with morphological differentiation, which is related to secondary metabolite production. In this study, we identified and characterized a diguanylate cyclase (DGC), CdgB, from Streptomyces diastatochromogenes 1628, which may be involved in c-di-GMP synthesis, through genetic and biochemical analyses. To further investigate the role of CdgB, the cdgB-deleted mutant strain Δ-cdgB and the cdgB-overexpressing mutant strain O-cdgB were constructed by genetic engineering. A phenotypic analysis revealed that the O-cdgB colonies exhibited reduced mycelium formation, whereas the Δ-cdgB colonies displayed wrinkled surfaces and shriveled mycelia. Notably, O-cdgB demonstrated a significant increase in the toyocamycin (TM) yield by 47.3%, from 253 to 374 mg/L, within 10 days. This increase was accompanied by a 6.7% elevation in the intracellular concentration of c-di-GMP and a higher transcriptional level of the toy cluster within four days. Conversely, Δ-cdgB showed a lower c-di-GMP concentration (reduced by 6.2%) in vivo and a reduced toyocamycin production (decreased by 28.9%, from 253 to 180 mg/L) after 10 days. In addition, S. diastatochromogenes 1628 exhibited a slightly higher inhibitory effect against Fusarium oxysporum f. sp. cucumerinum and Rhizoctonia solani compared to Δ-cdgB, but a lower inhibition rate than that of O-cdgB. The results imply that CdgB provides a foundational function for metabolism and the activation of secondary metabolism in S. diastatochromogenes 1628.

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Section: Strains Plasmids Primers and Culture Conditionsmentioning

confidence: 99%

“…The strain was grown at 28 • C in a glucose yeast mineral (GYM) medium [28] for a morphological analysis. Toyocamycin production was carried out using the same media and methods as previously reported [27]. Fusarium oxysporum f. sp.…”

Section: Strains Plasmids Primers and Culture Conditionsmentioning

confidence: 99%

CdgB Regulates Morphological Differentiation and Toyocamycin Production in Streptomyces diastatochromogenes 1628

Wang,

Zhang,

et al. 2024

IJMS

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“…Nevertheless, ribosomal A-sites for which charged cognate tRNA is sparse or absent can be occupied by uncharged cognate tRNA resulting in relA-mediated synthesis of the alarmone (p)ppGpp and the stringent response [ 36 , 37 ]. While certain other physiological conditions can induce ppGpp synthesis [ 37 ], it is a paradox that the known broad-ranging effects of (p)ppGpp on Streptomyces are at the later growth stages [ 38 , 39 ] (and also in Caulobacter [ 40 ]). The presence during the vegetative state of preformed UUA-cognate tRNA only needing the final base modification to be a substrate for aminoacylation is expected to facilitate a quick transition to the formation of functional acylated tRNA.…”

Section: Introductionmentioning

confidence: 99%

Streptomyces rare codon UUA: from features associated with 2 adpA related locations to candidate phage regulatory translational bypassing

Antonov,

O’Loughlin,

Gorohovski

et al. 2023

RNA Biology

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In Streptomyces species, the cell cycle involves a switch from an early and vegetative state to a later phase where secondary products including antibiotics are synthesized, aerial hyphae form and sporulation occurs. AdpA, which has two domains, activates the expression of numerous genes involved in the switch from the vegetative growth phase. The adpA mRNA of many Streptomyces species has a UUA codon in a linker region between 5’ sequence encoding one domain and 3’ sequence encoding its other and C-terminal domain. UUA codons are exceptionally rare in Streptomyces, and its functional cognate tRNA is not present in a fully modified and acylated form, in the early and vegetative phase of the cell cycle though it is aminoacylated later. Here, we report candidate recoding signals that may influence decoding of the linker region UUA. Additionally, a short ORF 5’ of the main ORF has been identified with a GUG at, or near, its 5’ end and an in-frame UUA near its 3’ end. The latter is commonly 5 nucleotides 5’ of the main ORF start. Ribosome profiling data show translation of that 5’ region. Ten years ago, UUA-mediated translational bypassing was proposed as a sensor by a Streptomyces phage of its host’s cell cycle stage and an effector of its lytic/lysogeny switch. We provide the first experimental evidence supportive of this proposal.

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Genetics of conditional extended mycelial cell viability of Streptomyces minutiscleroticus in deep starvation phase implicates the involvement of (p)ppGpp, clpX, and a histidine kinase sasA

Chatupale,

Pohnerkar

2024

Front. Microbiol.

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Bacterial lifespan ranges from a few hours to geological timescales. The prolonged survival trait under extreme energy starvation is essential for the perpetuation of their existence. The theme for long-term survival [long-term stationary phase (LTSP)] in the non-growing state may be dependent on the diversity in the environmental niche and the lifestyle of the bacteria, exemplified by longevity studies, albeit few, with model organisms. In the present study, we characterized the LTSP of mycelial cells of Streptomyces Minutiscleroticus, which remain metabolically active, demonstrate ongoing protein synthesis—killed by protein synthesis inhibitors—and remarkably by the cell-wall synthesis inhibitors, vancomycin, and ampicillin, suggesting “growth.” Their rapid turnover is also evident in ~10-fold loss of colony-forming unit (CFU) over a year, suggesting that for the death of one “old” cell, slightly less than one “new” cell is born. This longevity is consequent to (i) induction of the gene expression program effected by non-metabolizable, non-ionic osmolyte, sucrose, thus conditional, and (ii) possibly rendering this carbon utilizable by the production of a slow hydrolytic activity generating glucose, reinforcing the relevance of low-level energy resource for long term survival in the starvation phase. The viability parameters of LTSP cells measured through up to 90 days suggest that the stationary phase transitioning into LTSP following nutrient exhaustion is nearly quantitative. Expectedly, the viability in LTSP is (p)ppGpp/RelA dependent. Whereas mutation in chaperone clpX, negatively affects survival in stationary phase, overexpression of signal sensor-transducer histidine kinase, SasA8, enhances cell survivability. The relevance of longevity functions identified here requires further deduction of the genetic program.

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Physiology and Transcriptional Analysis of ppGpp-Related Regulatory Effects in Streptomyces diastatochromogenes 1628

Cited by 3 publications

References 29 publications

CdgB Regulates Morphological Differentiation and Toyocamycin Production in Streptomyces diastatochromogenes 1628

CdgB Regulates Morphological Differentiation and Toyocamycin Production in Streptomyces diastatochromogenes 1628

Streptomyces rare codon UUA: from features associated with 2 adpA related locations to candidate phage regulatory translational bypassing

Genetics of conditional extended mycelial cell viability of Streptomyces minutiscleroticus in deep starvation phase implicates the involvement of (p)ppGpp, clpX, and a histidine kinase sasA

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