Regulation of outer membrane biogenesis components LptD and Omp85 in cyanobacteria Anabaena sp. PCC 7120

• Applicant: Professor Dr. Enrico Schleiff
• Subject Area: Plant Physiology
• Term: from 2012 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 210961441

• Final Report Year: 2022

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
  
• (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
  
• (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
  
• (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
  
• (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
  
• (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
  
• (2015) The composition of the global and feature specific cyanobacterial core-genomes. Front Microbiol. 6:219
  Simm S, Keller M, Selymesi M, Schleiff E
  
• (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
  
• (2017) Die Verteilungsmechanismen von β-barrel Proteinen in Pflanzenzellen. BIOspektrum 23.127-129
  Klinger A, Ladig R, Schleiff E
  
• (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
  
• (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
  
• (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
  
• (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
  
• (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
  
• (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