title: Membrane targeting of MnSOD is essential for oxidative stress tolerance of nitrogen-fixing cultures of Anabaena sp. strain PCC7120.
abstract: The nitrogen-fixing cyanobacterium, Anabaena PCC7120 encodes for a membrane-targeted 30 kDa Mn-superoxide dismutase (MnSOD) and a cytosolic FeSOD. The MnSOD is post-translationally processed to 27 and 24 kDa forms in the cytosol and periplasm/thylakoid lumen. The extent of cleavage of signal and linker peptides at the N-terminus is dependent on the availability of combined nitrogen during growth. While the 24 and 27 kDa forms are present in near equal proportions under nitrogen-fixing conditions, the 24 kDa form is predominant under nitrogen-supplemented conditions. Individual contribution of these forms of MnSOD to total oxidative stress tolerance was analysed using recombinant Anabaena strains overexpressing either different molecular forms of MnSOD or MnSOD defective in the cleavage of signal/linker peptide. Targeting of MnSOD to the membrane and subsequent cleavage to release both the 24 and 27 kDa forms was essential for oxidative stress tolerance under nitrogen-fixing conditions. On the other hand, the cleavage of linker peptide was absolutely essential and the release of cytosolic 24 kDa form of MnSOD was obligatory for developing oxidative stress tolerance under nitrogen-supplemented conditions. Thus, a single MnSOD caters to the reduction of superoxide radical in both cytosol and thylakoid lumen/periplasm irrespective of the N-status of growth by regulating its cleavage. This is the first report on the physiological advantage of membrane-targeting and processing of MnSOD in either bacteria or plants. The higher oxidative stress tolerance offered by the cytosolic form of MnSOD has possibly resulted in retention of only the cytosolic form in bacterial non-nitrogen-fixers during evolution.
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, 400085, India.
authors.forename: Prashanth S
authors.initials: PS
authors.lastname: Raghavan
authors.forename: Hema
authors.initials: H
authors.lastname: Rajaram
authors.forename: Shree Kumar
authors.initials: SK
authors.lastname: Apte
mesh_terms: D017033:Anabaena / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D005798:Genes, Bacterial; D008970:Molecular Weight; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism; D009586:Nitrogen Fixation; D018384:Oxidative Stress; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D050505:Mutant Proteins; D011994:Recombinant Proteins; D013482:Superoxide Dismutase; D009584:Nitrogen