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Two enzymes together capable of cysteine biosynthesis are encoded on a cyanobacterial plasmid

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Abstract

The cyanobacterium Synechococcus sp. PCC 7942 contains two endogenous, genetically cryptic plasmids pANS and pANL, respectively. Characterization of the 3.8 kb Ba6 BamHI fragment of pANL identified three open reading frames which were transcriptionally regulated by sulfur availability and the protein CysR. One of these genes, designated srpG, encodes a protein which exhibits 67% amino acid identity to the Escherichia coli enzyme O-acetyl-l-serine (thiol)-lyase A. Overlapping the 3′ end of srpG is a second gene, designated srpH, which encodes a protein with similarity to the amino-terminal region of serine acetyltransferase enzymes. DNA hybridization results indicate that there is a second copy of srpG in Synechococcus sp. PCC 7942, which is consistent with previous isoenzyme studies on O-acetyl-l-serine (thiol)-lyase in cyanobacteria. The introduction of srpG and srpH into E. coli cysKcysM and cysE mutant strains, respectively, results in the complementation of the lesion in cysteine biosynthesis. Additionally, the E. coli cysK cysM strain containing srpG is able to utilize sulfate more efficiently than thiosulfate, indicating that SrpG is probably a type A O-acetyl-l-serine (thiol)-lyase. The possible function of these genes in the adaptation of cyanobacteria to sulfur stress is discussed.

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References

  • Allen MM (1968) Simple conditions for the growth of unicellular blue-green algae on plates. J Phycol 4: 1–4

    Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Molec Biol 215: 403–410

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1992) Current protocols in molecular biology. Green Publishing Associated and Wiley-Interscience, New York

    Google Scholar 

  • Becker MA, Kredich NM, Tomkins GM (1969) The purification and characterization of O-acetylserine sulfydrylase A from Salmonella typhimurium. J Biol Chem 244: 2418–2427

    Google Scholar 

  • Collier JL, Grossman AR (1992) Chlorosis induced by nutrient deprivation in Synechococcus sp. strain PCC 7942: Not all bleaching is the same. J Bacteriol 174: 4718–4726

    Google Scholar 

  • Denk D, Bock A (1987) l-cysteine biosynthesis in Escherichia coli: nucleotide sequence and expression of the serine acetyltransferase (cysE) gene from the wild-type and a cysteine excreting mutant. J Gen Microbiol 133: 515–525

    Google Scholar 

  • Diessner W, Schmidt A (1981) Isoenzymes of cysteine synthase in the cyanobacterium Synechococcus 6301. Z Pflanzenphysiol 102: 57–68

    Google Scholar 

  • Evans DJ, Jones R, Woodley PR, Wilborn JR, Robson RL (1991) Nucleotide sequence and genetic analysis of the Azotobacter chroococcum nifUSVWZM gene cluster, including a new gene (nifP) which encodes a serine acetyltransferase. J Bacteriol 173: 5457–5469

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13

    Google Scholar 

  • Filutowicz M, Waiter A, Hulanicka D (1982) Delayed inducibility of sulfite reductase in cysM mutants of S. typhimurium under anaerobic conditions. J Gen Microbiol 128: 1791–1794

    Google Scholar 

  • Green LS, Grossman AR (1988) Changes in sulfate transport characteristics and protein composition of Anacystis nidulans R2 during sulfur deprivation. J Bact 170: 583–587

    Google Scholar 

  • Green LS, Laudenbach DE, Grossman AR (1989) A region of a cyanobacterial genome required for sulfate transport. Proc Natl Acad Sci USA 86: 1949–1953

    Google Scholar 

  • Henikoff S (1987) Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol 155: 156–165

    Google Scholar 

  • Hulanicka MD, Kredich NM, Treiman DM (1974) The structural gene for O-acetylserine sulfhydrylase A in Salmonella typhimurium. Identity with the trzA locus. J Biol Chem 249: 867–872

    Google Scholar 

  • Hulanicka MD, Hallquist SG, Kredich NM, Mojica-AT (1979) Regulation of O-acetylserine sulfhydrylase B by l-cysteine in Salmonella typhimurium. J Bacteriol 140: 141–146

    Google Scholar 

  • Jensen TE, Rachlin JW (1984) Effect of varying sulfur deficiency on structural components of a cyanobacterium Synechococcus leopoliensis: a morphometric study. Cytobios 42: 35–46

    Google Scholar 

  • Kredich NM, Tomkins GM (1966) The enzymatic synthesis of l-cysteine in Escherichia coli and Salmonella typhimurium. J Biol Chem 244:4955–4965

    Google Scholar 

  • Kredich NM, Becker MA, Tomkins GM (1969) Purification and characterization of cysteine synthase, a bifunctional protein complex from Salmonella typhimurium. J Biol Chem 244: 2428–2439

    Google Scholar 

  • Lau RH, Doolittle WF (1979) Covalently closed circular DNA in closely related unicellular cyanobacteria. J Bacteriol 137: 648–652

    Google Scholar 

  • Lau RH, Sapienza C, Doolittle WF (1980) Cyanobacterial plasmids: their widespread occurrence, and the existence of regions of homology between plasmids in the same and different species. Molec Gen Genet 178: 203–211

    Google Scholar 

  • Laudenbach DE, Grossman AR (1991) Characterization and mutagenesis of sulfur-regulated genes in a cyanobacterium: evidence for function in sulfate transport. J Bacteriol 173: 2739–2750

    Google Scholar 

  • Laudenbach DE, Straus NA, Gendel S, Williams JP (1983) The large endogenous plasmid of Anacystis nidulans: mapping, cloning and localization of the origin of replication. Mol Gen Genet 192: 402–407

    Google Scholar 

  • Laudenbach DE, Reith ME, Straus NA (1988) Isolation, sequence analysis, and transcriptional studies of the flavodoxin gene from Anacystis nidulans R2. J Bacteriol 170: 258–265

    Google Scholar 

  • Laudenbach DE, Herbert SK, McDowell C, Fork DC, Grossman AG, Straus NA (1990) Cytochrome c-553 is not required for photosynthetic activity in the cyanobacterium Synechococcus. The Plant Cell 2: 913–924

    Google Scholar 

  • Laudenbach DE, Ehrhardt D, Green L, Grossman A (1991) Isolation and characterization of a sulfur-regulated gene encoding a periplasmically localized protein with sequence similarity to rhodanese. J Bacteriol 173: 2751–2760

    Google Scholar 

  • Lawry NH, Jensen TE (1979) Deposition of condensed phosphate as an effect of varying sulfur deficiency in the cyanobacterium Synechococcus sp. (Anacystis nidulans). Arch Microbiol 120: 1–7

    Google Scholar 

  • Leon J, Romero LC, Galvan F (1988) Intracellular levels and regulation of O-acetyl-l-serine sulthydrylase activity in Chlamydomonas reinhardtii. J Plant Physiol 132: 618–622

    Google Scholar 

  • Levy S, Danchin A (1988) Phylogeny of metabolic pathways: O-acetylserine sulphydrylase A is homologous to the tryptophan synthase beta subunit. Molec Microbiol 2: 777–783

    Google Scholar 

  • Nakamura T, Iwahashi H, Eguchi Y (1984) Enzymatic proof for the identity of the S-sulfocysteine synthase and cysteine synthase B of Salmonella typhimurium. J Bacteriol 158: 1122–1127

    Google Scholar 

  • Nakamura T, Kon Y, Iwahashi H, Eguchi Y (1983) Evidence that thiosulfate assimilation by Salmonella typhimurium is catalyzed by cysteine synthase B. J Bacteriol 156: 656–662

    Google Scholar 

  • Nalabolu SR, Tai C-H, Schnackerz KD, Cook PF (1992) Mechanism of O-acetylserine sulphydrylase from Salmonella typhimurium. Amino Acids 2: 119–125

    Google Scholar 

  • Passera C, Ghisi R (1982) ATP-sulphurylase and O-acetylserine sulphydrylase in isolated mesophyll protoplasts and bundle sheath strands of S-deprived maize leaves. J Exp Bot 33: 432–438

    Google Scholar 

  • Reith ME, Laudenbach DE, Straus NA (1986) Isolation and nucleotide sequence analysis of the ferredoxin I gene from the cyanobacterium Anacystis nidulans R2. J Bacteriol 168: 1319–1324

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger FS, Nicklen S, Coulson AR (1977) DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467

    Google Scholar 

  • Schmidt A, Erdle I, Kost HP (1982) Changes of C-phycocyanin in Synechococcus 6301 in relation to growth on various sulfur compounds. Z Naturforsch C37: 870–876

    Google Scholar 

  • Schmidt A (1990) Sulphur metabolism; cysteine synthase. In: Lea PJ (ed) Methods in plant biochemistry, vol. 3. Academic Press, London, pp 349–353

    Google Scholar 

  • Tabor S, Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci USA 84: 4767–4771

    Google Scholar 

  • Tandeau de Marsac N, Houmard J (1987) Advances in cyanobacterial molecular genetics. In: Fay P, Van Baalen C (eds) The cyanobacteria. Elsevier Science Publishing Company, New York, pp 251–302

    Google Scholar 

  • Tandeau de Marsac N, Borrias WE, Kuhlemeier CJ, Castetes AM, Van Arkel GA, van den Hondel CAAMJ (1982) A new approach for molecular cloning in cyanobacteria: cloning of an Anacystis nidulans met gene using a Tn901-induced mutant. Gene 20: 111–119

    Google Scholar 

  • Thomas PA (1983) Hybridization of denatured RNA transferred or dotted to nitrocellulose paper. Methods Enzymol 100: 255–266

    Google Scholar 

  • Utikilen HC, Heldal M, Knutsen G (1976) Characterization of sulfate uptake in Anacystis nidulans. Physiol Plant 34: 217–220

    Google Scholar 

  • Van den Hondel CAMJJ, Van Arkel GA (1980) Development of a cloning system in cyanobacteria. Antonie van Leeuwenhock 46: 228–229

    Google Scholar 

  • Van den Hondel CAMJJ, Keegstra W, Borrias WE, Van Arkel GA (1979) Homology of plasmids in strains of unicellular cyanobacteria. Plasmid 2: 323–333

    Google Scholar 

  • Wanner G, Henkelman A, Schmidt A, Kost HP (1986) Nitrogen and sulfur starvation of the cyanobacterium Synechococcus 6301. An ultra structural, morphometrical, and biochemical comparison. Z Naturforsch C41: 741–750

    Google Scholar 

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Communicated by H. Böhme

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Nicholson, M.L., Gaasenbeek, M. & Laudenbach, D.E. Two enzymes together capable of cysteine biosynthesis are encoded on a cyanobacterial plasmid. Molec. Gen. Genet. 247, 623–632 (1995). https://doi.org/10.1007/BF00290354

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