Oligoribonuclease is a central feature of cyclic diguanylate signaling in Pseudomonas aeruginosa
| Autor: | Undine Mechold, Jacquelyn D. Rich, Yafit Yarmiyhu, Ehud Banin, Denice C. Bay, Volkhard Kaever, Dorit Cohen, Joe J. Harrison, Matthew R. Parsek, Trevor E. Randall, Hadas Nevenzal |
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| Přispěvatelé: | Bar-Ilan University [Israël], Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), University of Calgary, University of Washington [Seattle], Hannover Medical School [Hannover] (MHH), E.B. was supported by Israel Science Foundation Grant 1124/12, D.C. was supported by a Federation of European Microbiological Societies fellowship, J.J.H. was supported by a Canada Research Chair from the Canadian Institutes of Health Research and Discovery Grant 435631 from the Natural Sciences and Engineering Research Council of Canada, and M.R.P. was supported by National Institute for Allergy and Infectious Disease Grant 2R01AI077628-05A1., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2015 |
| Předmět: |
MESH: Signal Transduction
[SDV]Life Sciences [q-bio] EAL domain MESH: Escherichia coli Proteins biofilm MESH: Reverse Transcriptase Polymerase Chain Reaction Homeostasis MESH: Cyclic GMP Cyclic GMP MESH: Bacterial Proteins MESH: Gene Expression Regulation Bacterial 0303 health sciences Multidisciplinary Reverse Transcriptase Polymerase Chain Reaction Escherichia coli Proteins MESH: Phosphorus-Oxygen Lyases Deoxyguanine Nucleotides Biological Sciences Biochemistry MESH: Homeostasis Pseudomonas aeruginosa MESH: Pseudomonas aeruginosa MESH: Exoribonucleases Second messenger system Phosphorus-Oxygen Lyases Intracellular Signal Transduction Exonuclease MESH: Mutation Blotting Western Biology MESH: Deoxyguanine Nucleotides 03 medical and health sciences Bacterial Proteins cyclic diguanylate MESH: Blotting Western 030304 developmental biology Nuclease Phosphoric Diester Hydrolases 030306 microbiology oligoribonuclease RNA Gene Expression Regulation Bacterial [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology Product inhibition Exoribonucleases Mutation Phosphodiester bond biology.protein MESH: Phosphoric Diester Hydrolases |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, 2015, 112 (36), pp.11359-64. ⟨10.1073/pnas.1421450112⟩ Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2015, 112 (36), pp.11359-64. ⟨10.1073/pnas.1421450112⟩ |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1421450112 |
| Popis: | International audience; The second messenger cyclic diguanylate (c-di-GMP) controls diverse cellular processes among bacteria. Diguanylate cyclases synthesize c-di-GMP, whereas it is degraded by c-di-GMP-specific phosphodiesterases (PDEs). Nearly 80% of these PDEs are predicted to depend on the catalytic function of glutamate-alanine-leucine (EAL) domains, which hydrolyze a single phosphodiester group in c-di-GMP to produce 5'-phosphoguanylyl-(3',5')-guanosine (pGpG). However, to degrade pGpG and prevent its accumulation, bacterial cells require an additional nuclease, the identity of which remains unknown. Here we identify oligoribonuclease (Orn)-a 3'→5' exonuclease highly conserved among Actinobacteria, Beta-, Delta- and Gammaproteobacteria-as the primary enzyme responsible for pGpG degradation in Pseudomonas aeruginosa cells. We found that a P. aeruginosa Δorn mutant had high intracellular c-di-GMP levels, causing this strain to overexpress extracellular polymers and overproduce biofilm. Although recombinant Orn degraded small RNAs in vitro, this enzyme had a proclivity for degrading RNA oligomers comprised of two to five nucleotides (nanoRNAs), including pGpG. Corresponding with this activity, Δorn cells possessed highly elevated pGpG levels. We found that pGpG reduced the rate of c-di-GMP degradation in cell lysates and inhibited the activity of EAL-dependent PDEs (PA2133, PvrR, and purified recombinant RocR) from P. aeruginosa. This pGpG-dependent inhibition was alleviated by the addition of Orn. These data suggest that elevated levels of pGpG exert product inhibition on EAL-dependent PDEs, thereby increasing intracellular c-di-GMP in Δorn cells. Thus, we propose that Orn provides homeostatic control of intracellular pGpG under native physiological conditions and that this activity is fundamental to c-di-GMP signal transduction. |
| Databáze: | OpenAIRE |
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