Regulation of outer membrane biogenesis components LptD and Omp85 in cyanobacteria Anabaena sp. PCC 7120
Final Report Abstract
In the course of the DFG funding we were able to establish a deep understanding of the function of the proteins involved in the biogenesis of the outer membrane of cyanobacteria including the two-partner secretion system, the β-barrel protein assembly machinery and the Lipid A transport machinery. The approach taken was a combinatory approach unifying the functional assays with structural determination either by crystallography (three structures), or by the analysis with methods like PELDOR or structural modelling. Especially the experimental approaches were challenging by the high protein demand and by the multitude of mutants required in case of PELDOR. We provided a hypothesis for the different functionality of the Omp85 proteins, which needs to be further challenged. The experimental work was complemented by bioinformatics, which assigned Omp85 proteins as core genes of the bacteria. Further, we gave insights into the functional cycle of the periplasmic holdase Tic22. The latter is one example for the bridging between the ancestral cyanobacterial system and the chloroplast system. In this line, we have defined the function of the LptD1 protein from Anabaena sp. Related to this machinery, we have identified and characterized the components of the LipidA transport system in Anabaena sp. The LptC protein discovered shows a remarkable extension when compared to E. coli LptC, nevertheless, the structural similarity clearly shows a structural conservation of this pathway. I would also like to mention that we had side observations like for anaCyp40, which were unexpected. After reprove our initial hypotheses, we continued the analysis of this protein with our established methodology to assign their function. We contributed with this study to the understaning of the regulation of the photosystems in cyanobacteria.
Publications
- (2012) Recycling and Tinkering- The evolution of protein transport to and into endosymbiotically derived organelles. In: Organelle Genetics: Evolution of organelle genomes and gene expression (Ed. E. C.E. Bullerwell), Springer Verlag Berlin, pp 175–216
Mirus O, Schleiff E
(See online at https://doi.org/10.1007/978-3-642-22380-8_8) - (2012) Structure and conservation of the periplasmic targeting factor Tic22 from plants and cyanobacteria. J Biol Chem. 287: 24164-24173
Tripp J, Hahn A, Koenig P, Flinner N, Bublak D, Brouwer EM, Ertel F, Mirus O, Sinning I, Tews I, Schleiff E
(See online at https://doi.org/10.1074/jbc.m112.341644) - (2014) Cell envelope components influencing filament length in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 196: 4026-4035
Burnat M, Schleiff E, Flores E
(See online at https://doi.org/10.1128/jb.02128-14) - (2014) Protein targeting and transport as a necessary consequence of increased cellular complexity. Cold Spring Harb Perspect Biol. 6(8). pii: a016055
Sommer MS, Schleiff E
(See online at https://doi.org/10.1101/cshperspect.a016055) - (2014) The cell envelope. In The Cell Biology of Cyanobacteria, (Eds. E. Flores and A. Herrero) Caister Academic Press, Norfolk, UK, pp 29-87
Hahn A, Schleiff E
(See online at https://doi.org/10.21775/9781910190739.03) - (2015) Functional properties of LptA and LptD in Anabaena sp. PCC 7120. Biol Chem. 396:1151-62
Hsueh YC, Brouwer EM, Marzi J, Mirus O, Schleiff E
(See online at https://doi.org/10.1515/hsz-2014-0322) - (2015) The composition of the global and feature specific cyanobacterial core-genomes. Front Microbiol. 6:219
Simm S, Keller M, Selymesi M, Schleiff E
(See online at https://doi.org/10.3389/fmicb.2015.00219) - (2016) Relative Orientation of POTRA Domains from Cyanobacterial Omp85 Studied by Pulsed EPR Spectroscopy. Biophys J. 110:2195-206
Dastvan R, Brouwer EM, Schuetz D, Mirus O, Schleiff E, Prisner TF
(See online at https://doi.org/10.1016/j.bpj.2016.04.030) - (2017) Die Verteilungsmechanismen von β-barrel Proteinen in Pflanzenzellen. BIOspektrum 23.127-129
Klinger A, Ladig R, Schleiff E
(See online at https://doi.org/10.1007/s12268-017-0774-8) - (2017) The cyanobacterial core-genome – global and specific features with a focus on secondary metabolites. In: Cyanobacteria: Omics and Manipulation (Ed. D.A. Los) Caister Academic Press. 1-34
Simm S, Schleiff E, Pernil R
(See online at https://doi.org/10.21775/9781910190555.01) - (2017) The plastid outer membrane localized LPTD1 is important for glycerolipid remodelling under phosphate starvation. Plant Cell Environ. 40:1643-1657
Hsueh YC, Ehmann C, Flinner N, Ladig R, Schleiff E
(See online at https://doi.org/10.1111/pce.12973) - (2019) Tic22 from Anabaena sp. PCC 7120 with holdase function involved in outer membrane protein biogenesis shuttles between plasma membrane and Omp85. Mol Microbiol. 111:1302-1316
Brouwer EM, Ngo G, Yadav S, Ladig R, Schleiff E
(See online at https://doi.org/10.1111/mmi.14222) - (2020) LptC from Anabaena sp. PCC 7120: Expression, purification and crystallization. Protein Expr Purif. 175:105689
Ngo G, Centola M, Krasnoselska G, Pogoryelov D, Yildiz Ö, Schleiff E
(See online at https://doi.org/10.1016/j.pep.2020.105689) - (2021) Comparative Phenotypic Analysis of Anabaena sp. PCC 7120 Mutants of Porin like Genes. J Microbiol Biotechnol. 31: 645-658
Schätzle H, Brouwer EM, Liebhart E, Stevanovic M, Schleiff E
(See online at https://doi.org/10.4014/jmb.2103.03009) - (2021) The Two TpsB-like Proteins in Anabaena sp. PCC 7120 Are Involved in Secretion of Selected Substrates. J Bacteriol. 203:e00568-20
Ngo G, Girbas M, Schätzle H, Hammer A, Safarian S, Hübinger M, Schleiff E
(See online at https://doi.org/10.1128/jb.00568-20)
