MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v06/MSV000092431/primaryOK2000000Roman Sobotkahttps://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=8b2872282c904fdbb5c1694ab961678bsobotka@alga.czMassIVE1,142UNIMOD:4 - "Iodoacetamide derivative."UNIMOD:35 - "Oxidation or Hydroxylation."ProteomicstimsTOF ProSynechocystis Sp. Pcc 6803 (ncbitaxon:1148)1Laboratory of Photosynthesis, Centre Algatech, Institute of Microbiology, The Czech Academy of Sciences, 37901 Trebon; Faculty of Science, University of South Bohemia, 37005, Ceske BudejoviceBiogenesis of the photosynthetic apparatus requires complicated molecular machinery, individual components of which are either poorly characterized or unknown. The BtpA protein has been described as a factor required for the stability of photosystem I (PSI) in cyanobacteria; however, how the BtpA stabilized PSI remains unexplained. To clarify the role of BtpA, we constructed and characterized the btpA-null mutant (ΔbtpA) in the cyanobacterium Synechocystis sp. PCC 6803. The mutant contained only c. 1% of chlorophyll and nearly no thylakoid membranes. However, this strain, growing only in the presence of glucose, was genetically unstable and readily generated suppressor mutations that restore the photoautotrophy. Two suppressor mutations were mapped into the hemA gene encoding glutamyl-tRNA reductase (GluTR) - the first enzyme of tetrapyrrole biosynthesis. Indeed, the GluTR was not detectable in the ΔbtpA mutant and the suppressor mutations restored biosynthesis of tetrapyrroles and photoautotrophy by increased GluTR expression or by improved GluTR stability/processivity. We further demonstrated that GluTR associates with a large BtpA oligomer and that BtpA is required for the stability of GluTR. Our results show that the BtpA protein is involved in the biogenesis of photosystems at the level of regulation of tetrapyrrole biosynthesis.A thylakoid biogenesis BtpA protein is required for the initial step of tetrapyrrole biosynthesis in cyanobacteria.Skotnicová Petra P, Srivastava Amit A, Aggarwal Divya D, Talbot Jana J, Karlínová Iva I, Moos Martin M, Mareš Jan J, Bučinská Lenka L, Koník Peter P, Šimek Petr P, Tichý Martin M, Sobotka Roman RCyanophyceae, GRP1/cytohesin 1, Thylakoid, protein complex, Proteins, Gene, stepk, cyanophytes, protein, Cyanobacteria, protein-containing complex, CG11633, blue-green algae, cytohesin/GRP1, protein polypeptide chains, Blue-Green, tetrapyrrole biosynthesis, native protein, natural protein, polypeptide chain, tetrapyrrole synthesis, Oxyphotobacteria, Blue-Green Bacteria, GRP1, Protein, Grp1, 3.1.3.48, Thylakoid Membranes, Gene Products, Cyanophycota, Blue Green Bacteria, tetrapyrrole formation, Cyanophyta, protein aggregate, l(2)SH0323, blue-green bacteria., CYH1, Membranes, Blue Green Algae, Step, CG11628, tetrapyrrole anabolism, PTPSTEP, Oxygenic photosynthetic bacteria, l(2)SH2 0323, proteins, Striatum-enriched protein-tyrosine phosphatase, photosynthetic membrane, cyanobacteria, Membrane, Bacteria, DmelCG11628, Protein Gene Products, Gene Proteins, Algae, Neural-specific protein-tyrosine phosphatase, STEP, l(2)k08110, Blue-Green Algae, Grana, Thylakoid Membrane, Blue Green, GPHMaterials, factor 8 deficiency, Glukose, 2-hydroxyethyl methacrylate, Mbp1, haemophilia A, Monohydrate, protein, Aphanocapsa sp. N-1, sci, (DL)-Isomer, Mutations, Membrane Tissues, protein polypeptide chains, Roles, congenital, Dextrose, HOW, How, Concepts, protein aggregate, myd, l(3)j5D5, Phyllobilins, multicellular organismal biosynthetic process, regulation of tetrapyrrole formation, 24B, classical hemophilia, increased, single-organism biosynthetic process, autosomal haemophilia a, tetrapyrrole anabolism, anabolism, Tissue, stru, Haemophilia A, E coli, Mbp-1, proteins, l(3)S053606, CG10293, FVIII, ecotype, DL-glucose, l(3)j5B5, glucose, Role Concepts, Synechocystis sp. (ATCC 27184), classic, Strains, 2-hydroxyethyl ester, Membrane Tissue, 0904/17, gyltl1b-b, glutamyl-tRNA reductase, results, regulation of tetrapyrrole synthesis., Blue-Green, tetrapyrrole biosynthesis, classic haemophilia, Role Concept, tetrapyrrole synthesis, SZ1, regulation of tetrapyrrole biosynthesis, MDDGA6, mKIAA0609, Role, Blue Green Bacteria, Genetic Materials, tetrapyrrole formation, cultivar, (alpha-D)-Isomer, Synechocystis sp. 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ATCC 27184, Glcapparatus, Materials, factor 8 deficiency, Glukose, 2-hydroxyethyl methacrylate, Mbp1, haemophilia A, Monohydrate, protein, Aphanocapsa sp. N-1, sci, (DL)-Isomer, Mutations, protein polypeptide chains, devices, Roles, congenital, Dextrose, HOW, How, Concepts, protein aggregate, myd, l(3)j5D5, Phyllobilins, multicellular organismal biosynthetic process, regulation of tetrapyrrole formation, 24B, classical hemophilia, single-organism biosynthetic process, appliances, autosomal haemophilia a, tetrapyrrole anabolism, anabolism, stru, Haemophilia A, Suppressor Mutations, E coli, Mbp-1, proteins, l(3)S053606, CG10293, FVIII, ecotype, DL-glucose, Genetic Suppressions, l(3)j5B5, glucose, Role Concepts, Synechocystis sp. 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(strain N-1), MDC1D, enr, enlarged, D Glucose, Material, a tetrapyrrole, Blue-Green Algae, Biocatalyst, anon-EST:Liang-2.39, Cistron, Grana, gtrA protein, Blue Green, Glucose Monohydrate, Strains and Sprains, AHF, gluco-hexose, hemophilia, big, Synechocystis 6803, bioformation, Glucose, Biocatalysts, P62, instruments, Gene, cyanophytes, Cyanobacteria, biosynthesis, protein-containing complex, froggy, Gyltl1a, blue-green algae, large, polypeptide chain, Oxyphotobacteria, 2-propenoic acid, Chlorophylls, Gene Products, regulation of tetrapyrrole anabolism, Cyanophyta, Sprains, classical haemophilia, l(3)s2612, Genetic, formation, MDDGB6, hemA protein, LARGE, cyanobacteria, Bacteria, synthesis, BPFD#36, Enzyme, Algae, Synechocystis sp. (strain PCC 6803), Historesin, great, DmelCG10293, HEMA, Thylakoid Membrane, tetrapyrroles, Anhydrous, glycol methacrylate, hemophilia A, Cyanophyceae, Synechocystis PCC6803, (beta-D)-Isomer, Tetrapyrrole, protein complex, clone 2.39, Thylakoid, Proteins, Sprain, qkr, l(3)S090417, Cistrons, Genetic Suppression, Concept, strain, Chlorophyll 740, DXS1253E, native protein, Photosystem I, equipment, natural protein, Blue-Green Bacteria, KH93F, Protein, Strain, Cyanophycota, Thylakoid Membranes, glu-tRNA reductase, 2-methyl-, Mutation, who, Membranes, Photosystem I Reaction Center, Oxygenic photosynthetic bacteria, Synechocystis sp. (PCC 6803), F8C, F8B, tetrapyrrole, Who/How, Membrane, Suppressor, Anhydrous Dextrose, Protein Gene Products, Gene Proteins, Synechocystis sp.PCC6803, qkr[93F], Synechocystis sp. ATCC 27184, GlcCyanophyceae, GRP1/cytohesin 1, Thylakoid, protein complex, Proteins, Gene, stepk, cyanophytes, protein, Cyanobacteria, protein-containing complex, CG11633, blue-green algae, cytohesin/GRP1, protein polypeptide chains, Blue-Green, tetrapyrrole biosynthesis, native protein, natural protein, polypeptide chain, tetrapyrrole synthesis, Oxyphotobacteria, Blue-Green Bacteria, GRP1, Protein, Grp1, 3.1.3.48, Thylakoid Membranes, Gene Products, Cyanophycota, Blue Green Bacteria, tetrapyrrole formation, Cyanophyta, protein aggregate, l(2)SH0323, blue-green bacteria., CYH1, Membranes, Blue Green Algae, Step, CG11628, tetrapyrrole anabolism, PTPSTEP, Oxygenic photosynthetic bacteria, l(2)SH2 0323, proteins, Striatum-enriched protein-tyrosine phosphatase, photosynthetic membrane, cyanobacteria, Membrane, Bacteria, DmelCG11628, Protein Gene Products, Gene Proteins, Algae, Neural-specific protein-tyrosine phosphatase, STEP, l(2)k08110, Blue-Green Algae, Grana, Thylakoid Membrane, Blue Green, GPH0PXD043762falseA thylakoid biogenesis BtpA protein is required for the initial step of tetrapyrrole biosynthesis in cyanobacteriaBiogenesis of the photosynthetic apparatus requires complicated molecular machinery, individual components of which are either poorly characterized or unknown. The BtpA protein has been described as a factor required for the stability of Photosystem I (PSI) in cyanobacteria; however, how the BtpA stabilized PSI remains unexplained. To clarify the role of BtpA, we constructed and characterized the btpA-null mutant (btpA) in the cyanobacterium Synechocystis sp. PCC 6803. The mutant contained only ~ 1% of chlorophyll and nearly no thylakoid membranes. However, this strain, growing only in the presence of glucose, was genetically unstable and readily generate suppressor mutations that restore the photoautotrophy. Two suppressor mutations were mapped into the hemA gene encoding glutamyl-tRNA reductase (GluTR) - the first enzyme of tetrapyrrole biosynthesis. Indeed, the GluTR was not detectable in the btpA mutant and the suppressor mutations restored biosynthesis of tetrapyrroles and photoautotrophy by increased GluTR expression or by improved GluTR stability/processivity. We further demonstrated that GluTR associates with a large BtpA oligomer and that BtpA is required for the stability of GluTR. Our results show that the BtpA protein is involved in the biogenesis of photosystems on the level of regulation of tetrapyrrole biosynthesis.Thu Jul 13 07:24:00 BST 2023MSV00009243137986097