title: Recruitment of a foreign quinone into the A(1) site of photosystem I. I. Genetic and physiological characterization of phylloquinone biosynthetic pathway mutants in Synechocystis sp. pcc 6803.
abstract: Genes encoding enzymes of the biosynthetic pathway leading to phylloquinone, the secondary electron acceptor of photosystem (PS) I, were identified in Synechocystis sp. PCC 6803 by comparison with genes encoding enzymes of the menaquinone biosynthetic pathway in Escherichia coli. Targeted inactivation of the menA and menB genes, which code for phytyl transferase and 1,4-dihydroxy-2-naphthoate synthase, respectively, prevented the synthesis of phylloquinone, thereby confirming the participation of these two gene products in the biosynthetic pathway. The menA and menB mutants grow photoautotrophically under low light conditions (20 microE m(-2) s(-1)), with doubling times twice that of the wild type, but they are unable to grow under high light conditions (120 microE m(-2) s(-1)). The menA and menB mutants grow photoheterotrophically on media supplemented with glucose under low light conditions, with doubling times similar to that of the wild type, but they are unable to grow under high light conditions unless atrazine is present to inhibit PS II activity. The level of active PS II per cell in the menA and menB mutant strains is identical to that of the wild type, but the level of active PS I is about 50-60% that of the wild type as assayed by low temperature fluorescence, P700 photoactivity, and electron transfer rates. PS I complexes isolated from the menA and menB mutant strains contain the full complement of polypeptides, show photoreduction of F(A) and F(B) at 15 K, and support 82-84% of the wild type rate of electron transfer from cytochrome c(6) to flavodoxin. HPLC analyses show high levels of plastoquinone-9 in PS I complexes from the menA and menB mutants but not from the wild type. We propose that in the absence of phylloquinone, PS I recruits plastoquinone-9 into the A(1) site, where it functions as an efficient cofactor in electron transfer from A(0) to the iron-sulfur clusters.
authors.affiliation: Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa 50011, USA.
authors.forename: T W
authors.initials: TW
authors.lastname: Johnson
authors.forename: G
authors.initials: G
authors.lastname: Shen
authors.forename: B
authors.initials: B
authors.lastname: Zybailov
authors.forename: D
authors.initials: D
authors.lastname: Kolling
authors.forename: R
authors.initials: R
authors.lastname: Reategui
authors.forename: S
authors.initials: S
authors.lastname: Beauparlant
authors.forename: I R
authors.initials: IR
authors.lastname: Vassiliev
authors.forename: D A
authors.initials: DA
authors.lastname: Bryant
authors.forename: A D
authors.initials: AD
authors.lastname: Jones
authors.forename: J H
authors.initials: JH
authors.lastname: Golbeck
authors.forename: P R
authors.initials: PR
authors.lastname: Chitnis
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics; D002734:Chlorophyll / Q000378:metabolism; D000458:Cyanobacteria / Q000235:genetics / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D029968:Escherichia coli Proteins; D005418:Flavodoxin / Q000378:metabolism; D005798:Genes, Bacterial; D006836:Hydro-Lyases / Q000235:genetics; D007425:Intracellular Membranes; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D008027:Light; D045342:Light-Harvesting Protein Complexes; D009154:Mutation; D010641:Phenotype; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D045331:Photosystem I Protein Complex; D010837:Vitamin K 1 / Q000096:biosynthesis*
chemical_list: D029968:Escherichia coli Proteins; D005418:Flavodoxin; D007506:Iron-Sulfur Proteins; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D002734:Chlorophyll; C001785:chlorophyll P 700; D010837:Vitamin K 1; D019883:Alkyl and Aryl Transferases; C412723:phytyltransferase; C112258:MenA protein, E coli; D006836:Hydro-Lyases; C021853:naphthoate synthetase