Site-directed mutagenesis of the CP47 protein of photosystem II: alteration of the basic residue 448R to 448G prevents the assembly of functional photosystem II centers under chloride-limiting conditions.
The psbB gene encodes the intrinsic chlorophyll protein CP47 (CPa-1), a component of photosystem II in higher plants, algae, and cyanobacteria. Oligonucleotide-directed mutagenesis has been used to introduce mutations into a segment of the psbB gene which encodes the large extrinsic loop E of CP47 in the cyanobacterium Synechocystis sp. PCC 6803. One mutation, R448G, produced a strain with impaired photosystem II activity. When grown in standard BG-11 media (480 microM chloride), this strain grew photoautotrophically at about 50% the rate of control strains and exhibited 63% of the control photosystem II activity. Quantum yield measurement at low light intensities indicated that this mutant had 55% of the fully functional photosystem II centers contained in control strains of Synechocystis. Upon exposure to high light intensities, the mutant strain exhibited a 2.2-fold increase in the rate of photoinactivation. When grown in BG-11 which was depleted in chloride (20 microM chloride), the mutant strain exhibited dramatically altered characteristics. Little or no growth was observed in the mutant while the control strains grew at nearly normal rates. Growth rates of the mutant strain could be restored by the addition of 480 microM bromide to the chloride-deficient BG-11 media. In the presence of glucose, the mutant and control strains grew at comparable rates under either chloride-sufficient or chloride-limiting conditions. Analysis of the mutant cell line grown in the absence of chloride and in the presence of glucose indicated that it exhibited essentially no capacity for oxygen evolution.(ABSTRACT TRUNCATED AT 250 WORDS)
Putnam-Evans
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Department of Biology, East Carolina University, Greenville, North Carolina 27858.
Bricker
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D001483:Base Sequence; D002712:Chlorides / Q000737:chemistry; D000458:Cyanobacteria / Q000235:genetics / Q000254:growth & development; D017931:DNA Primers / Q000737:chemistry; D005947:Glucose / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D045332:Photosystem II Protein Complex; D013329:Structure-Activity Relationship
D002712:Chlorides; D017931:DNA Primers; D045342:Light-Harvesting Protein Complexes; D046911:Macromolecular Substances; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; C070970:photosystem II, chlorophyll-binding protein, CP-47; D005947:Glucose; D010100:Oxygen