CC-4800 rbcL-V331A mt+ (45-3B) (non-pf2 progeny used for genetic analysis)
$30.00
From Robert J. Spreitzer, University of Nebraska, July 2014
Phenotype: requires acetate, sensitive to light
The 45-3B chloroplast mutant was recovered by screening a collection of acetate-requiring mutants for lack of complementation with a known Rubisco rbcL mutant (Spreitzer and Ogren 1983; Spreitzer et al. 1988). The mutant was crossed with pf2 mt- to confirm uniparental inheritance, and this mt+ progeny clone was analyzed. Mutant 45-3B results from a V331A substitution (GTT-GCT) in the chloroplast-encoded large subunit of Rubisco, which causes decreases in Rubisco CO2/O2 specificity and carboxylation catalytic efficiency (Chen and Spreitzer 1989). The V331A mutant-enzyme x-ray crystal structure has been solved (Karkehabadi et al. 2007). Selection for photosynthesis-competent revertants yielded true revertants and intragenic suppressors (see R40-9D and R61-2J) (Chen and Spreitzer 1989; Chen et al. 1991). Other types of revertants have been recovered (see R236-2A, R255-9A, and R258-3B) (Satagopan and Spreitzer, unpublished). This strain has been maintained with acetate medium in darkness since its isolation.
Chen Z, Spreitzer RJ (1989) Chloroplast intragenic suppression enhances the low CO2/O2 specificity of mutant ribulosebisphosphate carboxylase/oxygenase. J Biol Chem 264:3051-3053
Chen Z, Yu W, Lee JH, Diao R, Spreitzer RJ (1991) Complementing amino-acid substitutions within loop 6 of the alpha/beta-barrel active site influence the CO2/O2 specificity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase. Biochemistry 30:8846-8850
Karkehabadi S, Satagopan S, Taylor TC, Spreitzer RJ, Andersson I (2007) Structural analysis of altered large-subunit loop-6/carboxy-terminus interactions that influence catalytic efficiency and CO2/O2 specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase. Biochemistry 46:11080-11089
Spreitzer RJ, Al-Abed SR, Huether MJ (1988) Temperature-sensitive, photosynthesis-deficient mutants of Chlamydomonas reinhardtii. Plant Physiol 86:773-777
Spreitzer RJ, Ogren WL (1983) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297