Journal Article

Random Mutagenesis Targeted to the <i>psbA</i>II Gene of <i>Synechocystis</i> sp. PCC 6803 to Identify Functionally Important Residues in the D1 Protein of the Photosystem II Reaction Center

Akihiro Yamasato, Tomoe Kamada and Kimiyuki Satoh

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 5, pages 540-548
Published in print May 2002 | ISSN: 0032-0781
Published online May 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf066
Random Mutagenesis Targeted to the psbAII Gene of Synechocystis sp.  PCC 6803 to Identify Functionally Important Residues in the D1 Protein of the Photosystem II Reaction Center

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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–Ala357) 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 PSII. 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 PSII. A preliminary characterization of some of the mutants obtained in this study is provided.

Keywords: Key words: D1 protein — Photosystem II — psbA gene — Random mutagenesis — Synechocystis sp. PCC 6803.; Abbreviations: Mn-cluster, four-manganese cluster of the water oxidizing complex; P680, primary electron donor of photosystem II; Yz, secondary electron donor of photosystem II.

Journal Article.  5706 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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