Abstract
Anabaena sp. PCC 7120 mutants defective in phycobiliprotein biosynthesis or phycobilisome assembly were generated by transposon mutagenesis. Four mutants with grossly reduced content of the major phycobiliprotein, phycocyanin, were found to have insertions within the cpcBACDEFG1G2G3G4 operon coding for phycocyanin biosynthesis and assembly. The insertion in mutant B646 separated the promoter from the open reading frames and eliminated production of the phycocyanin α (CpcA) and β (CpcB) subunits. Insertion in cpcC in mutant B642 eliminated production of the L36 Rlinker polypeptide required for assembly of phycocyanin into the distal discs of the phycobilisome rod substructures. Mutants B64328 and B64407 had insertions, respectively, in cpcE and cpcF, genes coding for the subunits of the heterodimeric lyase which catalyzes the attachment of phycocyanobilin to the phycocyanin apo-α subunit. Mutant SB12, often unable to survive under low light, was found to have an insertion in the apcE gene coding for the large core-membrane linker (L128 CM) that provides the scaffold for assembly of the phycobilisome core. DNA sequencing 3′ of apcE revealed genes apcABC, coding for the α and β subunits of allophycocyanin and for the small core linker L7.8 C. Amino acid sequence comparisons showed that the ApcA and ApcB proteins are 37% identical and that each of these polypeptides is highly similar to corresponding polypeptides from the distantly related filamentous strains Calothrix sp. PCC7601 and Mastigocladus laminosus.
Similar content being viewed by others
References
Altschul SF, Giah W, Miller W, Myers EW and Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215: 403-410
Apt KE, Collier JL and Grossman AR (1995) Evolution of the phycobiliproteins. J Mol Biol 248: 79-96
Beale S (1993) Biosynthesis of phycobilins. Chem Rev 93: 139-216
Belknap W and Haselkorn R (1987) Cloning and light regulation of expression of the phycocyanin operon of the cyanobacterium Anabaena. EMBO J 6: 871-884
Berkelman TR and Lagarias JC (1986) Visualization of bilin-linked peptides and proteins in polyacrylamide gels. Anal Biochem 156: 194-201
Bhalerao RP and Gustafsson P (1994) Factors influencing the phycobilisome rod composition of the cyanobacterium Synechococcussp. PCC 7942: Effects of reduced phycocyanin content, lack of rod-linkers and over-expression of the rod-terminating linker. Physiol Plantarum 90: 187-197
Bhalerao RP, Collier JL, Gustafsson P and Grossman AR (1995) The structure of phycobilisomes in mutants of Synechococcussp. strain PCC 7942 devoid of specific linker polypeptides. Photochem Photobiol 61: 298-302
Brejc K, Ficner R Huber R, and Steinbacher S (1995) Isolation, crystallization, crystal structure analysis and refinement of allophycocyanin from the cyanobacterium Spirulina platensisat 2.3 α resolution. J. Mol Biol 249: 424-440
Bryant DA (1988) Genetic analysis of phycobilisome biosynthesis, assembly, structure, and function in the cyanobacterium Synechococcussp. PCC 7002. In: Stevens SE Jr and Bryant DA (eds) Light-Energy Transduction in Photosynthesis: Higher Plant and Bacterial Models, pp 62-90. The American Society of Plant Physiologists
Bryant DA (ed) (1994) The Molecular Biology of Cyanobacteria. Kluwer Academic Publishers, Dordrecht, the Netherlands
Bryant DA, Glazer AN and Eiserling FA (1976) Characterization and structural properties of the major biliproteins of Anabaenasp. Arch Microbiol 110: 61-75
Bryant DA, Stirewalt VL, Glauser M, Frank G, Sidler W and Zuber H (1991) A small multigene family encodes the rod-core linker polypeptides of Anabaenasp. PCC7120 phycobilisomes. Gene 107: 91-99
Cai Y and Wolk CP (1990) Use of a conditionally lethal gene in Anabaenasp. strain PCC 7120 to select for double recombinants and to entrap insertion sequences. J Bacteriol 172: 3138-3145
Curtis SE (1995) Unpublished data. GenBank accession number U21853.
DeLange RJ, Williams LC and Glazer AN (1981) The amino acid sequence of the β subunit of allophycocyanin. J Biol Chem 256: 9558-9566
de Lorimier R, Bryant DA, Porter RD, Liu W-Y, Jay E and Stevens SE Jr. (1984) Genes for the α and β subunits of phycocyanin. Proc Natl Acad Sci USA 81: 7946-7950
Ducret A, Sidler W, Wehrli E, Frank G and Zuber H (1996) Isolation, characterization and electron microscopy analysis of a hemidiscoidal phycobilisome type from the cyanobacterium Anabaenasp. PCC7120. Eur J Biochem 236: 1010-1024
Fairchild CD, Zhao J, Zhou J, Colson SE, Bryant DA and Glazer AN (1992) Phycocyanin α-subunit phycocyanobilin lyase. Proc Natl Acad Sci USA 89: 7017-7021
Glauser M, Bryant DA, Frank G, Wehrli E, Sidler W and Zuber H (1992) Phycobilisome structure in the cyanobacteria Mastigocladus laminosusand Anabaenasp. PCC7120: a new model. Eur J Biochem 205: 907-915
Glazer AN (1994) Adaptive variations in phycobilisome structure. In: Bittar EE and Barber J (eds) Advances in Molecular and Cell Biology, Vol 10, pp 119-149. JAI Press Inc., Greenwich, Connecticut
Glazer AN, Lundell DJ, Yamanaka G and Williams RC (1983) The structure of a ‘simple’ phycobilisome. Ann Microbiol (Inst. Pasteur) 134B: 159-180
Grossman AR, Schaefer MR, Chiang GG and Collier JL (1993) The phycobilisome, a light-harvesting complex responsive to environmental conditions. Microbiol Rev 57: 725-749
Hu NT, Thiel T, Giddings TH and Wolk CP (1981) New Anabaenaand Nostoccyanophages from sewage settling ponds. Virology 114: 236-246
Jung LJ, Chan CF and Glazer AN (1995) Candidate genes for the phycoerythrocyanin α subunit lyase. J Biol Chem 270: 12877-12884
Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, Kimura T, Hosouchi T, Matsuno A, Muraki A, Nakazaki N, Naruo K, Odumura S, Shimpo S, Takeuchi C, Wada T, Watanabe A, Yamada M, Yasuda M, and Tabata S (1996) Sequence analysis of the genome of the unicellular cyanobacterium Synechocystissp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res 3: 109-136
Klotz AV and Glazer AN (1987) γ-N-methylasparagine in phycobiliproteins. J Biol Chem 262: 17350-17355
Klotz AV, Thomas BA, Glazer AN and Blacher RW (1990) Detection of methylated asparagine and glutamine residues in polypeptides. Anal Biochem 186: 95-100
Kuritz T, Ernst A, Black TA and Wolk CP (1993) High resolution mapping of genetic loci of AnabaenaPCC7120 required for photosynthesis and nitrogen fixation. Mol Microbiol 8: 101-110
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685
Lundell DJ and Glazer AN (1983) Molecular architecture of a light-harvesting antenna. Structure of the 18S core-rod subassembly of the Synechococcus6301 phycobilisome. J Biol Chem 258: 894-901
Shestakov SV and Khuyen NT (1970) Evidence for genetic transformation in blue-green alga Anacystis nidulans. Mol Gen Genet 107: 372-375
Sidler WA (1994) Phycobilisome and phycobiliprotein structures. In: Bryant DA (ed) The Molecular Biology of Cyanobacteria, pp 139-216, Kluwer Academic Publishers, Dordrecht, the Netherlands
Swanson RV, de Lorimier R and Glazer AN (1992a) Genes encoding the phycobilisome rod substructure are clustered on the Anabaenachromosome: Characterization of the phycoerythrocyanin operon. J Bacteriol 174: 2640-2647
Swanson RV, Zhou J, Leary JA, Williams T, de Lorimier R, Bryant DA and Glazer AN (1992b) Characterization of phycocyanin produced by cpcEand cpcFmutants and identification of an intergenic suppressor of the defect in bilin attachment. J Biol Chem 267: 16146-16154
Wolk CP, Cai Y, Cardemil L, Flores E, Hohn B, Murry M, Schmetterer G, Schrautemeier B and Wilson R (1988) Isolation and complementation of mutants of Anabaenasp. strain PCC7120 unable to grow aerobically on dinitrogen. J Bacteriol 170: 1239-1244
Wolk CP, Cai Y and Panoff J-M (1991) Use of a transposon with luciferase as a reporter to identify environmentally responsive genes in a cyanobacterium. Proc Natl Acad Sci USA 88: 5355-5359
Wolk CP, Vonshak A, Kehoe P and Elhai J (1984) Construction of shuttle vectors capable of conjugative transfer from Escherichia colito nitrogen-fixing filamentous cyanobacteria. Proc Natl Acad Sci USA 81: 1561-1565
Yanisch-Perron C, Vieira J and Messing J (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33: 103-119
Yu M-H, Glazer AN and Williams RC (1981) Cyanobacterial phycobilisomes. Phycocyanin assembly in the rod substructures of Anabaena variabilisphycobilisomes. J Biol Chem 256: 13130-13136
Yu M-H and Glazer AN (1982) Cyanobacterial phycobilisomes. Role of the linker polypeptides in the assembly of phycocyanin. J Biol Chem 257: 3429-3433
Zhou J, Gasparich GE, Stirewalt VL, de Lorimier R and Bryant DA (1992) The cpcEand cpcFgenes of Synechococcussp. PCC7002: construction and phenotypic characterization of interposon mutants. J Biol Chem 267: 16138-16145
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cai, Y.A., Schwartz, S.H. & Glazer, A.N. Transposon insertion in genes coding for the biosynthesis of structural components of the Anabaena sp. phycobilisome. Photosynthesis Research 53, 109–120 (1997). https://doi.org/10.1023/A:1005816906539
Issue Date:
DOI: https://doi.org/10.1023/A:1005816906539