Abstract
We report on the sequencing and analysis of a 3,557-bp genomic DNA clone that is located between 4.8 and 1.2 kilobase pairs (kb) upstream of the rbcL gene and is capable of complementing a class of cyanobacterium Synechococcus sp. strain PCC7942 mutants requiring a high level of CO2. The upstream 2,704 bp of this sequence is novel, the remaining 852 bp having been reported by other workers. Four new open reading frames (ORFs) have been identified along with putative promoter elements. These ORFs, which could code for proteins of 7, 10.9, 11, and 58 kDa in size, have been named ORF 64, ccmK, ccmL, and ccmM, respectively. The last three have been named ccm genes on the basis that insertional mutagenesis of each produces a phenotype requiring a high level of CO2 (i.e., each produces a lesion in the CO2 concentrating mechanism). The putative gene product for the large ccmM ORF has three internally repeated regions and also has two possible DNA binding motifs. Two defined mutants in the 3,557-bp region, mutants PVU and P-N, have been more fully characterized. The PVU mutant has a drug marker inserted into the ccmL gene, and it possesses abnormal rod-shaped carboxysomes. The P-N mutant is a 2.64-kb deletion of DNA from the same position in ccmL to a region closer to rbcL. This mutant, which has previously been shown to lack carboxysomes and have soluble ribulosebiphosphate carboxylase/oxygenase activity, has now been shown to have a predominantly soluble carboxysomal carbonic anhydrase activity. Both mutants were found to possess carboxysomal carbonic anhydrase activities which are below wild-type levels, and in the P-N mutant this activity appears to be unstable. The results are discussed in terms of the possible interactions of putative ccm gene products in the process of carboxysome assembly and function.
References
34
Referenced
139
-
Badger M. R. 1987. The CO2 concentrating mechanism in aquatic phototrophs p. 219-274. In M. D. Hatch and N. K. Boardman (ed.) The biochemistry of plants: a comprehensive treatise vol 10. Photosynthesis. Academic Press New York.
(
10.1016/B978-0-12-675410-0.50010-5
) - Badger , M. R. , and A. Gallagher . 1987 . Adaptation of photosynthetic CO2 and HC03- accumulation by the cyanobacterium Synechococcus PCC6301 to growth at different inorganic carbon concentrations . Aust. J. Plant Physiol. 14 : 189 - 210 . / Aust. J. Plant Physiol. / Adaptation of photosynthetic CO2 and HC03- accumulation by the cyanobacterium Synechococcus PCC6301 to growth at different inorganic carbon concentrations by Badger M. R. (1987)
-
Badger , M. R. , and G. D. Price . 1989 . Carbonic anhydrase associated with the cyanobacterium Synechococcus PCC7942 . Plant Physiol. (Bethesda) 89 : 51 - 60 .
(
10.1104/pp.89.1.51
) / Plant Physiol. (Bethesda) / Carbonic anhydrase associated with the cyanobacterium Synechococcus PCC7942 by Badger M. R. (1989) -
Badger , M. R. , and G. D. Price . 1992 . The CO2 concentrating mechanism in cyanobacteria and microalgae . Physiol. Plant. 84 : 606 - 615 .
(
10.1111/j.1399-3054.1992.tb04711.x
) / Physiol. Plant. / The CO2 concentrating mechanism in cyanobacteria and microalgae by Badger M. R. (1992) -
Badger , M. R. , G. D. Price , and J. -W. Yu . 1991 . Selection and analysis of mutants of the CO2 concentrating mechanism in cyanobacteria . Can. J. Bot. 69 : 974 - 983 .
(
10.1139/b91-125
) / Can. J. Bot. / Selection and analysis of mutants of the CO2 concentrating mechanism in cyanobacteria by Badger M. R. (1991) -
Codd , G. A. , and W. J. N. Marsden . 1984 . The carboxysomes (polyhedral bodies) of autotrophic prokaryotes . Biol. Rev. 59 : 389 - 422 .
(
10.1111/j.1469-185X.1984.tb00710.x
) / Biol. Rev. / The carboxysomes (polyhedral bodies) of autotrophic prokaryotes by Codd G. A. (1984) -
Devereux , J. , P. Haeberli , and 0. Smithies . 1984 . A comprehensive set of sequence analysis programs for the VAX . Nucleic Acids Res. 12 : 387 - 395 .
(
10.1093/nar/12.1Part1.387
) / Nucleic Acids Res. / A comprehensive set of sequence analysis programs for the VAX by Devereux J. (1984) -
Dzelzkalns , V. A. , G. C. Owens , and L. Bogorad . 1984 . Chloroplast promoter driven expression of the chloramphenicol acetyl transferase gene in a cyanobacterium . Nucleic Acids Res. 12 : 8917 - 8925 .
(
10.1093/nar/12.23.8917
) / Nucleic Acids Res. / Chloroplast promoter driven expression of the chloramphenicol acetyl transferase gene in a cyanobacterium by Dzelzkalns V. A. (1984) -
Friedberg , D. , A. Kaplan , R. Ariel , M. Kessel , and J. SeUffers . 1989 . The 5'-flanking region of the gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase/oxygenase is crucial for growth of the cyanobacterium Synechococcus sp. strain PCC 7942 at the level of CO2 in air . J. Bacteriol. 171 : 6069 - 6076 .
(
10.1128/jb.171.11.6069-6076.1989
) / J. Bacteriol. / The 5'-flanking region of the gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase/oxygenase is crucial for growth of the cyanobacterium Synechococcus sp. strain PCC 7942 at the level of CO2 in air by Friedberg D. (1989) -
Fukuzawa , H. , E. Suzuki , Y. Komukai , and S. Miyachi . 1992 . A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon fixation in the cyanobacterium, Synechococcus PCC7942 . Proc. Natl. Acad. Sci. USA 89 : 4437 - 4441 .
(
10.1073/pnas.89.10.4437
) / Proc. Natl. Acad. Sci. USA / A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon fixation in the cyanobacterium, Synechococcus PCC7942 by Fukuzawa H. (1992) -
Golden , S. S. , J. Brusslan , and R. Haselkorn . 1987 . Genetic engineering of the cyanobacterial chromosome . Methods Enzymol. 153 : 215 - 231 .
(
10.1016/0076-6879(87)53055-5
) / Methods Enzymol. / Genetic engineering of the cyanobacterial chromosome by Golden S. S. (1987) -
Golden , S. S. , M. S. Nalty , and D. C. Cho . 1989 . Genetic relationship of two highly studied Synechococcus strains designated Anacystis nidulans . J. Bacteriol. 171 : 24 - 29 .
(
10.1128/jb.171.1.24-29.1989
) / J. Bacteriol. / Genetic relationship of two highly studied Synechococcus strains designated Anacystis nidulans by Golden S. S. (1989) -
Hawley , D. K. , and W. R. McClure . 1983 . Compilation and analysis of Escherichia coli promoter DNA sequences . Nucleic Acids Res 11 : 2237 - 2255 .
(
10.1093/nar/11.8.2237
) / Nucleic Acids Res / Compilation and analysis of Escherichia coli promoter DNA sequences by Hawley D. K. (1983) -
Knight , S. , I. Andersson , and C. I. Branden . 1990 . Crystallographic analysis of ribulose 1,5-biphosphate carboxylase from spinach at 2.4 A resolution: subunit interactions and active site . J. Mol. Biol. 215 : 113 - 160 .
(
10.1016/S0022-2836(05)80100-7
) / J. Mol. Biol. / Crystallographic analysis of ribulose 1,5-biphosphate carboxylase from spinach at 2.4 A resolution: subunit interactions and active site by Knight S. (1990) -
Lieman-Hurwitz , J. , R. Schwarz , F. Martinez , Z. Maor , L. Reinhold , and A. Kaplan . 1991 . Molecular analysis of high-CO2-requiring mutants indicates that genes in the region of rbc, including rbcS, are involved in the ability of cyanobacteria to grow under low CO2 . Can. J. Bot. 69 : 945 - 950 .
(
10.1139/b91-121
) / Can. J. Bot. / Molecular analysis of high-CO2-requiring mutants indicates that genes in the region of rbc, including rbcS, are involved in the ability of cyanobacteria to grow under low CO2 by Lieman-Hurwitz J. (1991) - Mead , D. A. , E. S. Skorupa , and B. Kemper . 1986 . Singlestranded DNA "blue " T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1 : 67 - 74 . / Protein Eng. / Singlestranded DNA "blue by Mead D. A. (1986)
-
Miller , A. G. , and D. T. Canvin . 1989 . Glycoaldehyde inhibits CO2 fixation in the cyanobacterium Synechococcus UTEX 625 without inhibiting the accumulation of inorganic carbon or the associated quenching of chlorophyll a fluorescence . Plant Physiol. 91 : 1044 - 1049 .
(
10.1104/pp.91.3.1044
) / Plant Physiol. / Glycoaldehyde inhibits CO2 fixation in the cyanobacterium Synechococcus UTEX 625 without inhibiting the accumulation of inorganic carbon or the associated quenching of chlorophyll a fluorescence by Miller A. G. (1989) -
Ogawa , T. 1992 . Identification and characterization of the ictA/ndhL gene product essential to inorganic carbon transport in Synechococcus PCC6803 . Plant Physiol. (Bethesda) 99 : 1604 - 1608 .
(
10.1104/pp.99.4.1604
) / Plant Physiol. (Bethesda) / Identification and characterization of the ictA/ndhL gene product essential to inorganic carbon transport in Synechococcus PCC6803 by Ogawa T. (1992) -
Pabo , C. O. , and R. T. Sauer . 1984 . Protein-DNA recognition. Annu . Rev. Biochem. 53 : 293 - 321 .
(
10.1146/annurev.bi.53.070184.001453
) / Rev. Biochem. / Protein-DNA recognition. Annu by Pabo C. O. (1984) - .Price G. D. Unpublished data.
-
Price , G. D. , and M. R. Badger . 1989 . Ethoxyzolamide inhibition of C02-dependent photosynthesis in the cyanobacterium Synechococcus PCC 7942 . Plant Physiol. (Bethesda) 89 : 44 - 50 .
(
10.1104/pp.89.1.44
) / Plant Physiol. (Bethesda) / Ethoxyzolamide inhibition of C02-dependent photosynthesis in the cyanobacterium Synechococcus PCC 7942 by Price G. D. (1989) -
Price , G. D. , and M. R. Badger . 1989 . Ethoxyzolamide inhibition of CO2 uptake in the cyanobacterium Synechococcus PCC7942 without apparent inhibition of internal carbonic anhydrase activity . Plant Physiol. (Bethesda) 89 : 37 - 43 .
(
10.1104/pp.89.1.37
) / Plant Physiol. (Bethesda) / Ethoxyzolamide inhibition of CO2 uptake in the cyanobacterium Synechococcus PCC7942 without apparent inhibition of internal carbonic anhydrase activity by Price G. D. (1989) -
Price , G. D. , and M. R. Badger . 1989 . Expression of human carbonic anhydrase in the cyanobacterium Synechococcus PCC7942 creates a high C02-requiring phenotype . Plant Physiol. (Bethesda) 91 : 505 - 513 .
(
10.1104/pp.91.2.505
) / Plant Physiol. (Bethesda) / Expression of human carbonic anhydrase in the cyanobacterium Synechococcus PCC7942 creates a high C02-requiring phenotype by Price G. D. (1989) -
Price , G. D. , and M. R. Badger . 1989 . Isolation and characterization of high-CO2 requiring mutants of the cyanobacterium Synechococcus PCC7942: two phenotypes that accumulate inorganic carbon but are unable to generate CO2 within the carboxysome . Plant Physiol. (Bethesda) 91 : 514 - 525 .
(
10.1104/pp.91.2.514
) / Plant Physiol. (Bethesda) / Isolation and characterization of high-CO2 requiring mutants of the cyanobacterium Synechococcus PCC7942: two phenotypes that accumulate inorganic carbon but are unable to generate CO2 within the carboxysome by Price G. D. (1989) -
Price , G. D. , and M. R. Badger . 1991 . Evidence for the role of carboxysomes in the cyanobacterial CO2 concentrating mechanism . Can. J. Bot. 69 : 963 - 973 .
(
10.1139/b91-124
) / Can. J. Bot. / Evidence for the role of carboxysomes in the cyanobacterial CO2 concentrating mechanism by Price G. D. (1991) -
Price , G. D. , J. R. Coleman , and M. R. Badger . 1992 . Association of carbonic anhydrase activity with carboxysomes isolated from the cyanobacterium, Synechococcus PCC7942 . Plant Physiol. (Bethesda) 100 : 784 - 793 .
(
10.1104/pp.100.2.784
) / Plant Physiol. (Bethesda) / Association of carbonic anhydrase activity with carboxysomes isolated from the cyanobacterium, Synechococcus PCC7942 by Price G. D. (1992) -
Reichelt , B. Y. , and S. F. Delaney . 1983 . The nucleotide sequence of the large subunit of ribulose-1,5-biphosphate carboxylase from the unicellular cyanobacterium Synechococcus PCC6301 . DNA 2 : 121 - 129 .
(
10.1089/dna.1983.2.121
) / DNA / The nucleotide sequence of the large subunit of ribulose-1,5-biphosphate carboxylase from the unicellular cyanobacterium Synechococcus PCC6301 by Reichelt B. Y. (1983) -
Reinhold , L. , R. Kosloff , and A. Kaplan . 1991 . A model for inorganic carbon fluxes and photosynthesis in cyanobacterial carboxysomes . Can. J. Bot. 69 : 984 - 988 .
(
10.1139/b91-126
) / Can. J. Bot. / A model for inorganic carbon fluxes and photosynthesis in cyanobacterial carboxysomes by Reinhold L. (1991) - Reinhold , L. , M. Zviman , and A. Kaplan . 1989 . A quantitative model for inorganic carbon fluxes and photosynthesis in cyanobacteria . Plant Physiol. Biochem. 27 : 945 - 954 . / Plant Physiol. Biochem. / A quantitative model for inorganic carbon fluxes and photosynthesis in cyanobacteria by Reinhold L. (1989)
-
Rippka R. J. B. Waterbury and R. Y. Stanier. 1981. Isolation and purification of cyanobacteria: some general principles p. 212-220. In M. P. Staff H. Stolp H. G. Truper A. Balows and H. G. Schlegel (ed.) The prokaryotes. Springer-Verlag Berlin.
(
10.1007/978-3-662-13187-9_8
) - Sambrook J. E. F. Fritsch and T. Maniatis. 1989. Molecular cloning: a laboratory manual 2nd ed. Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y.
-
Volokita , M. , D. Zenvirth , A. Kaplan , and L. Reinhold . 1984 . Nature of the inorganic carbon species actively taken up by the cyanobacterium, Anabaena variabifis . Plant Physiol. (Bethesda) 76 : 599 - 602 .
(
10.1104/pp.76.3.599
) / Plant Physiol. (Bethesda) / Nature of the inorganic carbon species actively taken up by the cyanobacterium, Anabaena variabifis by Volokita M. (1984) -
Wiatermans , J. F. G. M. , and A. de Mots . 1965 . Spectrophotometric characteristics of chlorophylls a and b and their pheophytins in ethanol . Biochim. Biophys. Acta 109 : 448 - 453 .
(
10.1016/0926-6585(65)90170-6
) / Biochim. Biophys. Acta / Spectrophotometric characteristics of chlorophylls a and b and their pheophytins in ethanol by Wiatermans J. F. G. M. (1965) -
Yu , J. - W. , G. D. Price , L. Song , and M. R. Badger . 1992 . Isolation of a putative carboxysomal carbonic anhydrase gene from the cyanobacterium, Synechococcus PCC7942 . Plant Physiol. (Bethesda) 100 : 794 - 800 .
(
10.1104/pp.100.2.794
) / Plant Physiol. (Bethesda) / Isolation of a putative carboxysomal carbonic anhydrase gene from the cyanobacterium, Synechococcus PCC7942 by Yu J. (1992)
Dates
Type | When |
---|---|
Created | 8 years, 9 months ago (Nov. 7, 2016, 12:58 p.m.) |
Deposited | 4 years, 1 month ago (July 29, 2021, 1:50 p.m.) |
Indexed | 4 weeks, 2 days ago (Aug. 7, 2025, 4:59 p.m.) |
Issued | 32 years, 4 months ago (May 1, 1993) |
Published | 32 years, 4 months ago (May 1, 1993) |
Published Print | 32 years, 4 months ago (May 1, 1993) |
@article{Price_1993, title={Analysis of a genomic DNA region from the cyanobacterium Synechococcus sp. strain PCC7942 involved in carboxysome assembly and function}, volume={175}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.175.10.2871-2879.1993}, DOI={10.1128/jb.175.10.2871-2879.1993}, number={10}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Price, G D and Howitt, S M and Harrison, K and Badger, M R}, year={1993}, month=may, pages={2871–2879} }