The necessity of multi-parameter normalization in cyanobacterial research: A case study of the PsbU in Synechocystis sp. PCC 6803 using CRISPRi.
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| Název: | The necessity of multi-parameter normalization in cyanobacterial research: A case study of the PsbU in Synechocystis sp. PCC 6803 using CRISPRi. |
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| Autoři: | Veit MC; BMBF Junior Research Group Biophotovoltaics, Department of Microbial Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Stauder R; Molecular Biology of Cyanobacteria Group, Department of Solar Materials Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Bai Y; BMBF Junior Research Group Biophotovoltaics, Department of Microbial Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Gabhrani R; BMBF Junior Research Group Biophotovoltaics, Department of Microbial Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Schmidt M; Centre for Chemical Microscopy (ProVIS), Department of Technical Biogeochemistry, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Klähn S; Molecular Biology of Cyanobacteria Group, Department of Solar Materials Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany., Lai B; BMBF Junior Research Group Biophotovoltaics, Department of Microbial Biotechnology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany. Electronic address: bin.lai@ufz.de. |
| Zdroj: | The Journal of biological chemistry [J Biol Chem] 2025 Nov; Vol. 301 (11), pp. 110763. Date of Electronic Publication: 2025 Sep 24. |
| Způsob vydávání: | Journal Article |
| Jazyk: | English |
| Informace o časopise: | Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology |
| Výrazy ze slovníku MeSH: | Synechocystis*/genetics , Synechocystis*/metabolism , Synechocystis*/growth & development , Photosystem II Protein Complex*/genetics , Photosystem II Protein Complex*/metabolism , Bacterial Proteins*/genetics , Bacterial Proteins*/metabolism , CRISPR-Cas Systems*, Oxygen/metabolism ; Photosynthesis ; Chlorophyll A/metabolism |
| Abstrakt: | Competing Interests: Conflict of interest The authors declare that they do not have any conflicts of interest with the content of this article. Photosystem II (PSII) is a multiprotein complex and plays a central role in oxygenic photosynthesis. PsbU, a 12 kDa subunit of PSII, is associated with thermotolerance and structural stabilization of the oxygen-evolving complex in cyanobacteria. Corresponding knockout strains showed decreased oxygen evolution rates, although the growth was not impaired. In this study, we provide further insights into the consequences of PsbU perturbations and propose to revisit the impact of PsbU on cell physiology. We made use of CRISPRi to knock down the psbU gene in Synechocystis sp. PCC 6803, and assessed previously described effects referred to different biomass parameters including optical density, chlorophyll a content and cell number. After knocking down psbU, the growth rate was decreased by 15% based on counting the cell number, while this effect was not observed when monitoring optical density. Furthermore, the oxygen evolution rate per cell in the psbU knockdown strain did not show a significant difference compared to the control groups, which was probably due to its larger cell size and higher chlorophyll a content per cell. The decreased quantum efficiency of pigments was compensated by the increased pigment content on the single-cell level in the knockdown strain. Our results complement previous analyses and highlight the importance of evaluating cyanobacterial physiology based on different biomass quantitative units to avoid misinterpretation of the results. (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Contributed Indexing: | Keywords: CRISPR; cell diameter; cell morphology; chlorophyll; cyanobacteria; gene interference; photosynthesis; photosystem II; polymerase chain reaction |
| Substance Nomenclature: | 0 (Photosystem II Protein Complex) 0 (Bacterial Proteins) S88TT14065 (Oxygen) YF5Q9EJC8Y (Chlorophyll A) |
| Entry Date(s): | Date Created: 20250926 Date Completed: 20251128 Latest Revision: 20251128 |
| Update Code: | 20251129 |
| PubMed Central ID: | PMC12590133 |
| DOI: | 10.1016/j.jbc.2025.110763 |
| PMID: | 41005478 |
| Databáze: | MEDLINE |
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Nájsť tento článok vo Web of Science | Abstrakt: | Competing Interests: Conflict of interest The authors declare that they do not have any conflicts of interest with the content of this article.<br />Photosystem II (PSII) is a multiprotein complex and plays a central role in oxygenic photosynthesis. PsbU, a 12 kDa subunit of PSII, is associated with thermotolerance and structural stabilization of the oxygen-evolving complex in cyanobacteria. Corresponding knockout strains showed decreased oxygen evolution rates, although the growth was not impaired. In this study, we provide further insights into the consequences of PsbU perturbations and propose to revisit the impact of PsbU on cell physiology. We made use of CRISPRi to knock down the psbU gene in Synechocystis sp. PCC 6803, and assessed previously described effects referred to different biomass parameters including optical density, chlorophyll a content and cell number. After knocking down psbU, the growth rate was decreased by 15% based on counting the cell number, while this effect was not observed when monitoring optical density. Furthermore, the oxygen evolution rate per cell in the psbU knockdown strain did not show a significant difference compared to the control groups, which was probably due to its larger cell size and higher chlorophyll a content per cell. The decreased quantum efficiency of pigments was compensated by the increased pigment content on the single-cell level in the knockdown strain. Our results complement previous analyses and highlight the importance of evaluating cyanobacterial physiology based on different biomass quantitative units to avoid misinterpretation of the results.<br /> (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.) |
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| ISSN: | 1083-351X |
| DOI: | 10.1016/j.jbc.2025.110763 |