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标题：RbcS suppressor mutations improve the thermal stability and CO2/O2 specificity of rbcL- mutant ribulose-1,5-bisphosphate carboxylase/oxygenase
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作者：Yu-Chun Du ; Seokjoo Hong ; Robert J. Spreitzer 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2000
卷号：97
期号：26
页码：14206-14211
DOI：10.1073/pnas.260503997
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：In the green alga Chlamydomonas reinhardtii, a Leu290-to-Phe (L290F) substitution in the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which is coded by the chloroplast rbcL gene, was previously found to be suppressed by second-site Ala222-to-Thr and Val262-to-Leu substitutions. These substitutions complement the photosynthesis deficiency of the L290F mutant by restoring the decreased thermal stability, catalytic efficiency, and CO2/O2 specificity of the mutant enzyme back to wild-type values. Because residues 222, 262, and 290 interact with the loop between {beta} strands A and B of the Rubisco small subunit, which is coded by RbcS1 and RbcS2 nuclear genes, it seemed possible that substitutions in this loop might also suppress L290F. A mutation in a nuclear gene, Rbc-1, was previously found to suppress the biochemical defects of the L290F enzyme at a posttranslational step, but the nature of this gene and its product remains unknown. In the present study, three nuclear-gene suppressors were found to be linked to each other but not to the Rbc-1 locus. DNA sequencing revealed that the RbcS2 genes of these suppressor strains have mutations that cause either Asn54-to-Ser or Ala57-to-Val substitutions in the small-subunit {beta}A/{beta}B loop. When present in otherwise wild-type cells, with or without the resident RbcS1 gene, the mutant small subunits improve the thermal stability of wild-type Rubisco. These results indicate that the {beta}A/{beta}B loop, which is unique to eukaryotic Rubisco, contributes to holoenzyme thermal stability, catalytic efficiency, and CO2/O2 specificity. The small subunit may be a fruitful target for engineering improved Rubisco.