| Popis: |
Genes encoding enzymes of the biosynthetic pathway leading to phylloquinone, the secondary electron acceptor of photosystem (PS) I, were identified inSynechocystis sp. PCC 6803 by comparison with genes encoding enzymes of the menaquinone biosynthetic pathway inEscherichia coli. Targeted inactivation of themenA 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 μE m−2 s−1), with doubling times twice that of the wild type, but they are unable to grow under high light conditions (120 μE m−2 s−1). The menA andmenB 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 menAand 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 themenA and menB mutant strains contain the full complement of polypeptides, show photoreduction of FA and FB 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 A1site, where it functions as an efficient cofactor in electron transfer from A0 to the iron-sulfur clusters. |
