{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T08:04:30Z","timestamp":1774166670708,"version":"3.50.1"},"reference-count":39,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T00:00:00Z","timestamp":1639440000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/130478\/2017"],"award-info":[{"award-number":["SFRH\/BD\/130478\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["IF\/00256\/2015"],"award-info":[{"award-number":["IF\/00256\/2015"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["PTDC\/BIA-OUT\/29540\/2017"],"award-info":[{"award-number":["PTDC\/BIA-OUT\/29540\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Cyanobacteria are a group of photosynthetic prokaryotes that contribute to primary production on a global scale. These microorganisms release vesicles to the extracellular environment, spherical nanosized structures, derived essentially from the outer membrane. Even though earlier works in model Gram-negative bacteria have hypothesized that outer membrane stability is crucial in vesicle formation, the mechanisms determining vesicle biogenesis in cyanobacteria remain unknown. Here, we report on the identification of six candidate genes encoding outer membrane proteins harboring SLH\/OprB-domains in the genome of the model cyanobacterium Synechocystis sp. PCC 6803. Using a genetics-based approach, one gene was found to encode an essential protein (Slr1841), while the remaining five are not essential for growth under standard conditions. Vesicle production was monitored, and it was found that a mutant in the gene encoding the second most abundant SLH\/OprB protein in Synechocystis sp. PCC 6803 outer membrane (Slr1908) produces more vesicles than any of the other tested strains. Moreover, the Slr1908-protein was also found to be important for iron uptake. Altogether, our results suggest that proteins containing the SLH\/OprB-domains may have dual biological role, related to micronutrient uptake and to outer membrane stability, which, together or alone, seem to be involved in cyanobacterial vesicle biogenesis.<\/jats:p>","DOI":"10.3390\/plants10122757","type":"journal-article","created":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T22:06:10Z","timestamp":1639519570000},"page":"2757","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["The Role of Outer Membrane Protein(s) Harboring SLH\/OprB-Domains in Extracellular Vesicles\u2019 Production in Synechocystis sp. PCC 6803"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7732-5717","authenticated-orcid":false,"given":"Delfim","family":"Cardoso","sequence":"first","affiliation":[{"name":"MABBS\u2014Mestrado em Aplica\u00e7\u00f5es em Biotecnologia e Biologia Sint\u00e9tica, Faculdade de Ci\u00eancias, Universidade do Porto, R. Campo Alegre s\/n, 4169-007 Porto, Portugal"},{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2881-9125","authenticated-orcid":false,"given":"Steeve","family":"Lima","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"MCbiology Doctoral Program, ICBAS\u2014Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar, Universidade do Porto, R. Jorge de Viterbo Ferreira 228, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4012-0069","authenticated-orcid":false,"given":"Jorge","family":"Matinha-Cardoso","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4396-2122","authenticated-orcid":false,"given":"Paula","family":"Tamagnini","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"Departamento de Biologia, Faculdade de Ci\u00eancias, Universidade do Porto, R. Campo Alegre s\/n, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0938-152X","authenticated-orcid":false,"given":"Paulo","family":"Oliveira","sequence":"additional","affiliation":[{"name":"i3S\u2014Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"IBMC\u2014Instituto de Biologia Molecular e Celular, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"Departamento de Biologia, Faculdade de Ci\u00eancias, Universidade do Porto, R. Campo Alegre s\/n, 4169-007 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1042\/BST20180252","article-title":"Answers to naysayers regarding microbial extracellular vesicles","volume":"47","author":"Coelho","year":"2019","journal-title":"Biochem. Soc. Trans."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1083\/jcb.201211138","article-title":"Extracellular vesicles: Exosomes, microvesicles, and friends","volume":"200","author":"Raposo","year":"2013","journal-title":"J. Cell Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/nrm.2017.125","article-title":"Shedding light on the cell biology of extracellular vesicles","volume":"19","author":"Raposo","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1146\/annurev.micro.091208.073413","article-title":"Biological Functions and Biogenesis of Secreted Bacterial Outer Membrane Vesicles","volume":"64","author":"Kulp","year":"2010","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1038\/nrmicro3525","article-title":"Outer-membrane vesicles from Gram-negative bacteria: Biogenesis and functions","volume":"13","author":"Schwechheimer","year":"2015","journal-title":"Nat. Rev. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1074\/jbc.M110.185744","article-title":"Selective Sorting of Cargo Proteins into Bacterial Membrane Vesicles","volume":"286","author":"Haurat","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.cub.2013.10.077","article-title":"An Ecological Network of Polysaccharide Utilization among Human Intestinal Symbionts","volume":"24","author":"Coyne","year":"2014","journal-title":"Curr. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lima, S., Matinha-Cardoso, J., Tamagnini, P., and Oliveira, P. (2020). Extracellular Vesicles: An Overlooked Secretion System in Cyano-bacteria. Life, 10.","DOI":"10.3390\/life10080129"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.tim.2016.03.005","article-title":"Bacterial Vesicle Secretion and the Evolutionary Origin of the Eukaryotic Endomembrane System","volume":"24","author":"Gould","year":"2016","journal-title":"Trends Microbiol."},{"key":"ref_10","first-page":"427","article-title":"Endosymbiotic origin and differential loss of eukaryotic genes","volume":"524","author":"Ku","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9824","DOI":"10.1073\/pnas.1307701110","article-title":"Present and future global distributions of the marine Cyanobacteria Prochlorococcus and Synechococcus","volume":"110","author":"Flombaum","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1126\/science.1243457","article-title":"Bacterial Vesicles in Marine Ecosystems","volume":"343","author":"Biller","year":"2014","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1128\/JB.182.5.1191-1199.2000","article-title":"Cyanobacterial Cell Walls: News from an Unusual Prokaryotic Envelope","volume":"182","author":"Hoiczyk","year":"2000","journal-title":"J. Bacteriol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"586","DOI":"10.1074\/mcp.M300137-MCP200","article-title":"Isolation of Outer Membrane of Synechocystis sp. PCC 6803 and Its Proteomic Characterization","volume":"3","author":"Huang","year":"2004","journal-title":"Mol. Cell. Proteom."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"20198","DOI":"10.1074\/jbc.M116.746131","article-title":"Outer Membrane Proteins Derived from Non-cyanobacterial Lineage Cover the Pepti-doglycan of Cyanophora paradoxa Cyanelles and Serve as a Cyanelle Diffusion Channel","volume":"291","author":"Kojima","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00371-17","DOI":"10.1128\/JB.00371-17","article-title":"Outer Membrane Permeability of Cyanobacterium Synechocystis sp. Strain PCC 6803: Studies of Passive Diffusion of Small Organic Nutrients Reveal the Absence of Classical Porins and Intrinsically Low Permeability","volume":"199","author":"Kowata","year":"2017","journal-title":"J. Bacteriol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"645","DOI":"10.4014\/jmb.2103.03009","article-title":"Comparative Phenotypic Analysis of Anabaena sp. PCC 7120 Mutants of Porin-like Genes","volume":"31","author":"Brouwer","year":"2021","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1093\/dnares\/dsu018","article-title":"Comparative Analysis of the Primary Transcriptome of Synechocystis sp. PCC 6803","volume":"21","author":"Kopf","year":"2014","journal-title":"DNA Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1111\/1462-2920.15324","article-title":"A unique porin meditates iron-selective transport through cyanobacterial outer membranes","volume":"23","author":"Qiu","year":"2021","journal-title":"Environ. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2779","DOI":"10.1128\/JB.183.9.2779-2784.2001","article-title":"Genes Essential to Iron Transport in the Cyanobacterium Synechocystis sp. Strain PCC 6803","volume":"183","author":"Katoh","year":"2001","journal-title":"J. Bacteriol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1146\/annurev.micro.112408.134247","article-title":"TonB-Dependent Transporters: Regulation, Structure, and Function","volume":"64","author":"Noinaj","year":"2010","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"01512","DOI":"10.1128\/AEM.01512-18","article-title":"Outer Membrane Iron Uptake Pathways in the Model Cyanobacterium Synechocystis sp. Strain PCC 6803","volume":"84","author":"Qiu","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"12520","DOI":"10.1074\/jbc.M709907200","article-title":"FutA2 Is a Ferric Binding Protein from Synechocystis PCC 6803","volume":"283","author":"Badarau","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1074\/mcp.M200043-MCP200","article-title":"Proteomics of Synechocystis sp. Strain PCC 6803","volume":"267","author":"Huang","year":"2002","journal-title":"Mol. Cell. Proteom."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3293","DOI":"10.1099\/00221287-148-10-3293","article-title":"Localization and function of the IdiA homologue Slr1295 in the cyanobacterium Synechocystis sp. strain PCC 6803","volume":"148","author":"Michel","year":"2002","journal-title":"Microbiology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"22168","DOI":"10.1038\/srep22168","article-title":"The Transcriptional Landscape of the Photosynthetic Model Cyanobacterium Synechocystis sp. PCC 6803","volume":"6","author":"Semeniuk","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"12424","DOI":"10.1038\/srep12424","article-title":"[NiFe]-hydrogenase is essential for cyanobacterium Synechocystis sp. PCC 6803 aerobic growth in the dark","volume":"5","author":"Checchetto","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2354","DOI":"10.1111\/1462-2920.14095","article-title":"Identification of inner membrane translocase components of TolC-mediated secretion in the cyanobacterium Synechocystis sp. PCC 6803","volume":"20","author":"Pacheco","year":"2018","journal-title":"Environ. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1111\/1462-2920.13172","article-title":"The versatile TolC-like Slr1270 in the cyanobacterium Synechocystis sp. PCC 6803","volume":"18","author":"Oliveira","year":"2016","journal-title":"Environ. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1093\/femsle\/fnv163","article-title":"Detection of outer membrane vesicles in Synechocystis PCC 6803","volume":"362","author":"Pardo","year":"2015","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1250","DOI":"10.1128\/JB.01090-13","article-title":"Role for Myco-bacterium tuberculosis Membrane Vesicles in Iron Acquisition","volume":"196","author":"Casadevall","year":"2014","journal-title":"J. Bacteriol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4473","DOI":"10.1093\/emboj\/19.17.4473","article-title":"Bacterial SLH domain proteins are non-covalently anchored to the cell surface via a conserved mechanism involving wall polysaccharide pyruvylation","volume":"19","author":"Mesnage","year":"2000","journal-title":"EMBO J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1128\/br.35.2.171-205.1971","article-title":"Purification and properties of unicellular blue-green algae (order Chroococcales)","volume":"35","author":"Stanier","year":"1971","journal-title":"Bacteriol. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"D493","DOI":"10.1093\/nar\/gkx922","article-title":"20 years of the SMART protein domain annotation resource","volume":"46","author":"Letunic","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"D458","DOI":"10.1093\/nar\/gkaa937","article-title":"SMART: Recent updates, new developments and status in 2020","volume":"49","author":"Letunic","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1128\/jcm.24.3.330-332.1986","article-title":"Rapid microprocedure for isolating detergent-insoluble outer membrane proteins from Haemophilus species","volume":"24","author":"Carlone","year":"1986","journal-title":"J. Clin. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Fisher, M.L., Allen, R., Luo, Y., and Curtiss, R. (2013). Export of Extracellular Polysaccharides Modulates Adherence of the Cyano-bacterium Synechocystis. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0074514"},{"key":"ref_38","first-page":"539","article-title":"Chapter Twenty-Four -Synthetic Biology in Cyanobacteria: Engineering and Analyzing Novel Functions","volume":"Volume 497","author":"Voigt","year":"2011","journal-title":"Methods in Enzymology"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1631","DOI":"10.1111\/1462-2920.12600","article-title":"HesF, an exoprotein required for filament adhesion and aggregation in Anabaena sp. PCC 7120","volume":"17","author":"Oliveira","year":"2015","journal-title":"Environ. Microbiol."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/10\/12\/2757\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:47:31Z","timestamp":1760168851000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/10\/12\/2757"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,14]]},"references-count":39,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["plants10122757"],"URL":"https:\/\/doi.org\/10.3390\/plants10122757","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,14]]}}}