Structural and biochemical analysis of a phosin from<i>Streptomyces chartreusis</i>reveals a combined polyphosphate‐ and metal‐binding fold

Werten, Sebastiaan; Rustmeier, N.H.; Gemmer, Maximilian; Hinrichs, W.

doi:10.1002/1873-3468.13476

Cited by 9 publications

(18 citation statements)

References 56 publications

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“…We observed that in all Streptomyces species, the pap–pitH1 operon is linked, and in opposite orientation, to the pstSCAB cluster and the ppk gene encoding the polyphosphate kinase ( Figure 2 ), a fact that has not been reported previously. In addition, the gene SCO4144, downstream of ppk , encodes a protein with 67% identity to the phosin PptA that binds metals and polyphosphate [ 81 ]. The protein encoded by SCO4144 contains all of the histidine residues characteristics of metal-binding proteins and the arginine residues for sulphate binding found in PptA.…”

Section: The Pap-pith1-pstscab-ppta-nuda-ppk Sumentioning

confidence: 99%

Molecular Mechanisms of Phosphate Sensing, Transport and Signalling in Streptomyces and Related Actinobacteria

Martı́n

Liras

2021

IJMS

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Phosphorous, in the form of phosphate, is a key element in the nutrition of all living beings. In nature, it is present in the form of phosphate salts, organophosphates, and phosphonates. Bacteria transport inorganic phosphate by the high affinity phosphate transport system PstSCAB, and the low affinity PitH transporters. The PstSCAB system consists of four components. PstS is the phosphate binding protein and discriminates between arsenate and phosphate. In the Streptomyces species, the PstS protein, attached to the outer side of the cell membrane, is glycosylated and released as a soluble protein that lacks its phosphate binding ability. Transport of phosphate by the PstSCAB system is drastically regulated by the inorganic phosphate concentration and mediated by binding of phosphorylated PhoP to the promoter of the PstSCAB operon. In Mycobacterium smegmatis, an additional high affinity transport system, PhnCDE, is also under PhoP regulation. Additionally, Streptomyces have a duplicated low affinity phosphate transport system encoded by the pitH1–pitH2 genes. In this system phosphate is transported as a metal-phosphate complex in simport with protons. Expression of pitH2, but not that of pitH1 in Streptomyces coelicolor, is regulated by PhoP. Interestingly, in many Streptomyces species, three gene clusters pitH1–pstSCAB–ppk (for a polyphosphate kinase), are linked in a supercluster formed by nine genes related to phosphate metabolism. Glycerol-3-phosphate may be transported by the actinobacteria Corynebacterium glutamicum that contains a ugp gene cluster for glycerol-3-P uptake, but the ugp cluster is not present in Streptomyces genomes. Sugar phosphates and nucleotides are used as phosphate source by the Streptomyces species, but there is no evidence of the uhp gene involved in the transport of sugar phosphates. Sugar phosphates and nucleotides are dephosphorylated by extracellular phosphatases and nucleotidases. An isolated uhpT gene for a hexose phosphate antiporter is present in several pathogenic corynebacteria, such as Corynebacterium diphtheriae, but not in non-pathogenic ones. Phosphonates are molecules that contains phosphate linked covalently to a carbon atom through a very stable C–P bond. Their utilization requires the phnCDE genes for phosphonates/phosphate transport and genes for degradation, including those for the subunits of the C–P lyase. Strains of the Arthrobacter and Streptomyces genera were reported to degrade simple phosphonates, but bioinformatic analysis reveals that whole sets of genes for putative phosphonate degradation are present only in three Arthrobacter species and a few Streptomyces species. Genes encoding the C–P lyase subunits occur in several Streptomyces species associated with plant roots or with mangroves, but not in the laboratory model Streptomyces species; however, the phnCDE genes that encode phosphonates/phosphate transport systems are frequent in Streptomyces species, suggesting that these genes, in the absence of C–P lyase genes, might be used as surrogate phosphate transporters. In summary, Streptomyces and related actinobacteria seem to be less versatile in phosphate transport systems than Enterobacteria.

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Section: The Pap-pith1-pstscab-ppta-nuda-ppk Sumentioning

confidence: 99%

Molecular Mechanisms of Phosphate Sensing, Transport and Signalling in Streptomyces and Related Actinobacteria

Martı́n

Liras

2021

IJMS

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“…On its side, SLI_4383/PptA is described as a 388 AA long protein bearing a TTA leucine codon, possible target for bldA regulation [ 21 ] and possessing a conserved histidine alpha-helical domain (“CHAD” domain), considered as a polyphosphate binding module [ 19 , 20 ]. Interestingly, a comparison of the predicted protein product of SLI_4383 / pptA (GenBank entry EFD67904.1) to putative orthologues from other Streptomyces species revealed the presence of N-terminal extensions of various length whose extremity is highly conserved in almost all orthologues but which is missing in the protein of S. lividans as it is annotated at the Sanger center ( Figure 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…In the recently determined crystal structure of S. chartreusis PptA [ 20 ], this N terminal region comprises part of the first α-helix of the protein fold. Since in the S. lividans genome (GenBank entry GG657756.1) a highly similar sequence is present immediately upstream of the designated pptA translational start codon, it seems likely that this codon was erroneously assigned to an internal GTG (Val) codon.…”

Section: Resultsmentioning

confidence: 99%

“…This indicated that these mutants suffered from phosphate stress/low phosphate availability. The phosin PptA possesses a CHAD domain forming a positively charged tunnel thought to accommodate negatively charged polyP [ 19 , 20 ]. This polyP binding protein is thus most likely involved in the degradation of polyP to supply phosphate intracellularly.…”

Section: Discussionmentioning

confidence: 99%

“…Proteins with CHAD domain are usually found associated with polyP granules in various organisms and are called phosins (Tumlirsch and Jendrossek 2017; Lorenzo-Orts, et al, 2019). The CHAD domain is thought to constitute a positively charged tunnel accommodating negatively charged polyP [ 18 , 19 , 20 ]. SLI_4382 encodes a protein of the nudix hydrolase family bearing similarities with enzymes involved in the cleavage of diadenosine hexaphosphate (Ap6A) into two molecules of ATP.…”

Section: Introductionmentioning

confidence: 99%

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The Phosin PptA Plays a Negative Role in the Regulation of Antibiotic Production in Streptomyces lividans

et al. 2021

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In Streptomyces, antibiotic biosynthesis is triggered in phosphate limitation that is usually correlated with energetic stress. Polyphosphates constitute an important reservoir of phosphate and energy and a better understanding of their role in the regulation of antibiotic biosynthesis is of crucial importance. We previously characterized a gene, SLI_4384/ppk, encoding a polyphosphate kinase, whose disruption greatly enhanced the weak antibiotic production of Streptomyces lividans. In the condition of energetic stress, Ppk utilizes polyP as phosphate and energy donor, to generate ATP from ADP. In this paper, we established that ppk is co-transcribed with its two downstream genes, SLI_4383, encoding a phosin called PptA possessing a CHAD domain constituting a polyphosphate binding module and SLI_4382 encoding a nudix hydrolase. The expression of the ppk/pptA/SLI_4382 operon was shown to be under the positive control of the two-component system PhoR/PhoP and thus mainly expressed in condition of phosphate limitation. However, pptA and SLI_4382 can also be transcribed alone from their own promoter. The deletion of pptA resulted into earlier and stronger actinorhodin production and lower lipid content than the disruption of ppk, whereas the deletion of SLI_4382 had no obvious phenotypical consequences. The disruption of ppk was shown to have a polar effect on the expression of pptA, suggesting that the phenotype of the ppk mutant might be linked, at least in part, to the weak expression of pptA in this strain. Interestingly, the expression of phoR/phoP and that of the genes of the pho regulon involved in phosphate supply or saving were strongly up-regulated in pptA and ppk mutants, revealing that both mutants suffer from phosphate stress. Considering the presence of a polyphosphate binding module in PptA, but absence of similarities between PptA and known exo-polyphosphatases, we proposed that PptA constitutes an accessory factor for exopolyphosphatases or general phosphatases involved in the degradation of polyphosphates into phosphate.

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Polyphosphate Granules and Acidocalcisomes

Jendrossek

2020

Bacterial Organelles and Organelle-Like Inclusions

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Structural and biochemical analysis of a phosin fromStreptomyces chartreusisreveals a combined polyphosphate‐ and metal‐binding fold

Cited by 9 publications

References 56 publications

Molecular Mechanisms of Phosphate Sensing, Transport and Signalling in Streptomyces and Related Actinobacteria

Molecular Mechanisms of Phosphate Sensing, Transport and Signalling in Streptomyces and Related Actinobacteria

The Phosin PptA Plays a Negative Role in the Regulation of Antibiotic Production in Streptomyces lividans

Polyphosphate Granules and Acidocalcisomes

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