A novel family of nonribosomal peptides modulate collective behavior in<i>Pseudovibrio</i>bacteria isolated from marine sponges

Ióca, Laura P.; Dai, Yitao; Kunakom, Sylvia; Diaz‐Espinosa, Jennifer; Krunić, Aleksej; Crnkovic, Camila M.; Orjala, Jimmy; Sanchez, Laura M.; Ferreira, Antonio G.; Berlinck, Roberto G. S.; Eustáquio, Alessandra S.

doi:10.1101/2020.12.15.422899

Cited by 3 publications

(23 citation statements)

References 61 publications

(98 reference statements)

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“…In contrast, the ∆pppD mutant produced only PA in agreement with previous results with pppD::neo (Figs. 2, S9-S11) (10).…”

Section: Resultsmentioning

confidence: 99%

“…1; Table S1). NRPS pppA::neo mutants helped us identify the products of the ppp BGC, which we termed pseudovibriamides A, and B (PA and PB) (10). P. brasiliensis Ab134 also accumulates a third product, pseudovibriamide C (PC), detected here by mass spectrometry analysis.…”

Section: Introductionmentioning

confidence: 82%

“…For instance, motility, mediated by bacterial metabolites, is known to be important for host colonization (9). We previously identified a link between swarming motility and pseudovibriamides (10). The ppp gene cluster that encodes pseudovibriamides (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…We have previously reported a link between a biosynthetic gene cluster (BGC) in Pseudovibrio brasiliensis Ab134 and its swarming motility (10). Strain Ab134 was isolated from marine sponge Arenosclera brasiliensis (11).…”

Section: Introductionmentioning

confidence: 99%

“…Strain Ab134 was isolated from marine sponge Arenosclera brasiliensis (11). The BGC, which we termed Pseudovibrio and Pseudomonas non-ribosomal peptide (ppp), is present in 2/3 of sequenced Pseudovibrio genomes, and sporadically found in Pseudomonas γ-Proteobacteria known to interact with terrestrial plants and insects (10). Moreover, a ppp-like BGC was recently reported from Microbulbifer γ-Proteobacteria likewise isolated from marine sponges (12).…”

Section: Introductionmentioning

confidence: 99%

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Pseudovibriamides fromPseudovibriomarine sponge bacteria promote swarming motility via transcriptional modulation

Dai,

Lourenzon,

Ióca

et al. 2024

Preprint

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Pseudovibrioα-Proteobacteria have been repeatedly isolated from marine sponges and proposed to be beneficial to the host. Bacterial motility is known to contribute to host colonization. We have previously identified pseudovibriamides A and B, produced in culture byPseudovibrio brasiliensisAb134, and shown that pseudovibriamide A promotes flagellar motility. Pseudovibriamides are encoded in a hybrid nonribosomal peptide synthetase-polyketide synthase gene cluster that also includes several accessory genes. Pseudovibriamide A is a linear heptapeptide and pseudovibriamide B is a nonadepsipeptide derived from pseudovibriamide A. Here we define the borders of the pseudovibriamides gene cluster, assign function to biosynthetic genes using reverse genetics and test the hypothesis that pseudovibriamides impact motility by modulating gene transcription. RNA-seq transcriptomic analyses of strains having different compositions of pseudovibriamides suggested that both pseudovibriamides A and B affect genes potentially involved in motility, and that a compensatory mechanism is at play in mutants that produce only pseudovibriamide A, resulting in comparable swarming motility as the wild type. The data gathered suggest that pseudovibriamides A and B have opposite roles in modulating a subset of genes, with pseudovibriamide B having a primary effect in gene activation, and pseudovibriamide A on inhibition. Finally, we observed many differentially expressed genes (up to 29% of the total gene number) indicating that pseudovibriamides have a global effect on transcription that goes beyond motility.ImportanceMarine sponges are found throughout the oceans from tropical coral reefs to polar sea floors, playing crucial roles in marine ecosystems.Pseudovibriobacteria have been proposed to contribute to sponge health. We have previously shown that pseudovibriamides produced byPseudovibrio brasiliensispromote bacterial motility, a behavior that is beneficial to bacterial survival and to host colonization. The gene cluster that encodes pseudovibriamide biosynthesis is found in two thirds ofPseudovibriogenomes. This gene cluster is also present inPseudomonasbacteria that interact with terrestrial plants and animals. Here we first assign function to pseudovibriamide biosynthetic genes using reverse genetics. We then show that pseudovibriamides play a major role in transcriptional regulation, affecting up to 29% ofP. brasiliensisgenes, including motility genes. Thus, this work gives insights into pseudovibriamide biosynthesis and provides evidence that they are signaling molecules relevant to bacterial motility and to other yet to be identified phenotypes.

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“…In contrast, the ∆pppD mutant produced only PA in agreement with previous results with pppD::neo (Figs. 2, S9-S11) (10).…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Pseudovibriamides fromPseudovibriomarine sponge bacteria promote swarming motility via transcriptional modulation

Dai,

Lourenzon,

Ióca

et al. 2024

Preprint

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Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

Dai,

Pita,

Eustáquio

2024

Preprint

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The filter feeding capacity of marine sponges contributes to biogeochemical cycling and they are also involved in habitat formation, properties that are critical to marine ecology. Sponge-associated microbes are crucial to the functional roles provided by sponges. α-Proteobacteria belonging to thePseudovibriogenus have been isolated from many different marine sponge genera and have been proposed to contribute to sponge health. We recently reported specialized metabolites we named pseudovibriamides fromPseudovibrio brasiliensisAb134. The pseudovibriamide encodingpppgene cluster is found in two thirds ofPseudovibriogenomes. Pseudovibriamides coordinate motility and biofilm formation, behaviors that are known to be important for host colonization. Although reverse genetics methods to delete genes via homologous recombination have been established, no self-replicative vectors have been reported forPseudovibrio. We show that plasmid vectors containing three different broad-host-range replicons, RSF1010, RK2, and pBBR1, can be used inP. brasiliensisfor fluorescent protein expression and consequent labeling. We then applied GFP and mCherry expressing strains to answer the question of whether pseudovibriamides affect the uptake ofP. brasiliensisbyAplysina aerophobasponges.P. brasiliensiscell counts decreased in the sponge aquaria at an equivalent rate for wild-type and pseudovibriamide-defective ΔpppAmutant strains, indicating that the sponge filters each strain indiscriminately under the conditions tested. Yet, the filtering capacity varied for each sponge individual tested, stressing the importance of performing experiments with wild-type and mutant bacterial strains in the same aquarium to allow for rigorous conclusions, which is now enabled with the methods established here.ImportanceMarine sponges are ecosystem engineers. They transform nutrients into a bioavailable form for other marine organisms. Microbes are critical to the functional roles provided by sponges because they expand the metabolic capabilities of the sponge host. Yet, most of our knowledge on sponge microbes comes from genomic studies, since cultivability and the ability to perform genetics with sponge bacterial isolates is limited. The genusPseudovibrioof α-Proteobacteria has consistently been isolated from marine sponges and it has been hypothesized to contribute to marine sponge health. Moreover,Pseudovibriobacteria are a source of antibiotics and other secondary metabolites with the potential to be developed into pharmaceuticals. Here we established vectors for the expression of fluorescent proteins inPseudovibriobacteria and demonstrated their utility inin vivostudies with marine sponges. The availability of genetic tools is important to enable us to explore the emerging ecological and biotechnological potential ofPseudovibriobacteria.

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Formation of the Ureido Bond in Anabaenopeptins by ApnA from Planktothrix agardhii CYA 126/8.

Philmus,

Hemscheidt

2024

Preprint

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The process leading to the formation of the ureido bond linking lysine and tyrosine/arginine, respectively, in the anabaenopeptins has been reconstituted in vitro. Trapping experiments suggest that an enzyme bound lysine -carbamic acid is an intermediate. Labeling experiments with 18O-bicarbonate suggest that a carbamic acid mixed anhydride is formed from the tyrosine/arginine thioester and subsequently rearranged to yield the ureido-dipeptide.

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A novel family of nonribosomal peptides modulate collective behavior inPseudovibriobacteria isolated from marine sponges

Cited by 3 publications

References 61 publications

Pseudovibriamides fromPseudovibriomarine sponge bacteria promote swarming motility via transcriptional modulation

Pseudovibriamides fromPseudovibriomarine sponge bacteria promote swarming motility via transcriptional modulation

Evaluation of vectors for gene expression inPseudovibriobacteria and their application inAplysinamarine sponge studies

Formation of the Ureido Bond in Anabaenopeptins by ApnA from Planktothrix agardhii CYA 126/8.

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