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. 2020 Feb 5;10:1894. doi: 10.1038/s41598-020-58726-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Slr7083 and HmpF_Syn are involved in twitching motility in Synechocystis. (A) Merged GFP fluorescence and chlorophyll autofluorescence (red) and bright field micrographs of Synechocystis cells expressing, Slr7083-GFP, HmpF_Syn-YFP or PilB-GFP from P_cpc560 (Slr7083) or P_trc (HmpF_Syn, PilB). Expression of PilB-GFP in PCC-M resulted in the same localization pattern (data not shown). White triangles indicate focal spots and crescent-like formations. Scale bars: 5 μm. (B) Merged bright field and chlorophyll autofluorescence micrographs of motile and non-motile Synechocystis WT, Δslr7083 and ΔhmpF_Syn mutant cells. Below, motility tests of three single colonies from indicated cells streaked on BG11 plates and illuminated from only one direction are shown. (C) Growth curve of Synechocystis WT, Δslr7083 and ΔhmpF_Syn mutant strains grown in quadruples at standard growth conditions. OD₇₅₀ values were recorded once a day for 15 d. Error bars show the standard deviation (n = 4). (D) Excerpt of interacting proteins of interest from mass spectrometry analysis of anti-GFP co-immunoprecipitations of Synechocystis cells expressing HmpF_Syn-YFP from P_trc. (E) Beta-galactosidase assays of E. coli cells co-expressing indicated translational fusion constructs of all possible pair-wise combinations of Slr7083 with HmpF_Syn grown for 1 d at 30 °C. Quantity values are given in Miller Units per milligram LacZ of the mean results from three independent colonies. Error bars indicate standard deviations (n = 3). Neg: pKNT25 plasmid carrying hmpF_Syn co-transformed with empty pUT18C. Pos: Zip/Zip control. Values indicated with * are significantly different from the negative control. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Dunnett’s multiple comparison test and one-way ANOVA).