Journal
PLANT AND CELL PHYSIOLOGY
Volume 43, Issue 5, Pages 540-548Publisher
OXFORD UNIV PRESS
DOI: 10.1093/pcp/pcf066
Keywords
D1 protein; photosystem II; psbA gene; random mutagenesis; Synechocystis sp PCC 6803
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More than one hundred mutants of Synechocystis sp. PCC 6803 impaired in photoautotrophic growth were generated by in vitro random PCR mutagenesis targeted to a region of the psbAII gene corresponding to a 210 amino acid (Ser148-Ala057) segment of the D1 protein. The 90 random mutants that could translate the full-length D1 protein carried 1-9 (on average 3.0) amino acid substitutions in the targeted region. Mutations were often found in the obligate photoheterotrophic strains at specific residues that have been reported or speculated to be important in the function of PSII, such as Y161, H198, H272, E333 and H337. This verifies the usefulness of the present method to Identify functionally important residues in PSIL Other residues that were often mutated in the strains with impaired photoautotrophy included non-charged residues around the lumenal edges of transmembrane helices C, D and E, such as I192 and N296. Eleven mutants carried a single-point mutation in residues, such as Q165, Q187, W278, A294 and N298, and these identified the functional importance of these residues, most of which were on the donor side of PSIL A preliminary characterization of some of the mutants obtained in this study is provided.
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