Abstract
Many herbicides that inhibit photosynthesis in plants also inhibit photosynthesis in bacteria. We have isolated three mutants of the photosynthetic bacterium Rhodobacter sphaeroides that were selected for increased resistance to the herbicide terbutryne. All three mutants also showed increased resistance to the known electron transfer inhibitor o-phenanthroline. The primary structures of the mutants were determined by recombinant DNA techniques. All mutations were located on the gene coding for the L-subunit resulting in these changes Ile229 → Met, Ser223 → Pro and Tyr222 → Gly. The mutations of Ser223 is analogous to the mutation of Ser264 in the D1 subunit of photosystem II in green plants, strengthening the functional analogy between D1 and the bacterial L-subunit. The changed amino acids of the mutant strains form part of the binding pocket for the secondary quinone, Q b . This is consistent with the idea that the herbicides are competitive inhibitors for the Q b binding site. The reaction centers of the mutants were characterized with respect to electron transfer rates, inhibition constants of terbutryne and o-phenanthroline, and binding constants of the quinone UQ0 and the inhibitors. By correlating these results with the three-dimensional structure obtained from x-ray analysis by Allen et al. (1987a, 1987b), the likely positions of o-phenanthroline and terbutryne were deduced. These correspond to the positions deduced by Michel et al. (1986a) for Rhodopseudomonas viridis.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- ATP:
-
adenosine 5′-triphosphate
- Bchl:
-
bacteriochlorophyll
- Bphe:
-
bacteriopheophytin
- bp:
-
basepair
- cyt c2+ :
-
reduced form of cytochrome c
- DEAE:
-
diethylami-noethyl
- EDTA:
-
ethylenediamine tetraacetic acid
- Fe2+ :
-
non-heme iron atom
- LDAO:
-
lauryl dimethylamine oxide
- Pipes:
-
piperazine-N,N′-bis-2-ethane-sulfonic acid
- PSII:
-
photosystem II
- RC:
-
reaction center
- SDS:
-
sodium dodecylsulfate
- Tris:
-
tris(hydroxy-methyl)aminomethane
- UQ0 :
-
2,3-dimethoxy-5-methyl benzoquinone
- UQ10 :
-
ubiquinone 50
References
Allen JP, Feher G, Yeates TO, Komiya H and Rees DC (1987a) Structure of the reaction center from Rhodobacter sphaeroides R26: The cofactors. Proc Natl Acad Sci USA 84: 5730–5734.
Allen JP, Feher G, Yeates TO, Komiya H and Rees DC (1987b) Structure of the reaction center from Rhodobacter sphaeroides R26: The protein subunits. Proc Natl Acad Sci USA 84: 6162–6166.
Barber J (1987) Rethinking the structure of the photosystem two reaction center. Trends Biochem Sci 12: 123–124.
Birnboim HC and Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acid Res 7: 1513–1523.
Brown AE, Gilbert CW, Guy R and Arntzen CJ (1984) Triazine herbicide resistance in the photosynthetic bacterium Rhodopseudomonas sphaeroides. Proc Natl Acad Sci USA 81: 6310–6314.
Brudvig GW, Worland ST and Sauer K (1983) Procedure for rapid isolation of photosynthetic reaction centers using cytochrome c affinity chromatography. Proc Natl Acad Sci USA 80: 683–686.
Bylina EJ and Youvan DC (1987) Genetic engineering of herbicide resistance: saturation mutagenesis of isoleucine 229 of the reaction center L subunit. Z Naturforsch 42c: 769–774.
Cogdell RJ, Monger TG and Parson WW (1975) Carotenoid triplet states in reaction centers from Rhodopseudomonas sphaeroides and Rhodospirillum rubrum. Biochim Biophys Acta 408: 189–199.
Crouse GF, Fritschauf A and Lehrach H (1983) An integrated and simplified approach to cloning into plasmids and single-stranded phages. Meth Enz 101: 78–89.
DeVitry C and Diner BA (1984) Photoaffinity labeling of the azidoatrazine receptor site in reaction centers of Rhodopseudomonas sphaeroides. FEBS Lett 167: 327–331.
Diner BA, Schenck CC and deVitry C (1984) Effect of inhibitors, redox state and isoprenoid chain length on the affinity of ubiquinone for the secondary acceptor binding site in the reaction centers of photosynthetic bacteria. Biochim Biophys Acta 766: 9–20.
Erickson JM, Rahire M, Bennoun P, Delepelaire P, Diner BA and Rochaix JD (1984) Herbicide resistance in Chlamydomonas reinhardtii results from a mutation in the chloroplast gene for the 32-kilodalton protein of photosystem II. Proc Natl Acad Sci USA 81: 3617–3621.
Erickson JM and Rochaix JD (1985) In: Galan GA (ed.) Abstracts. First International Congress of Plant Molecular Biology, p 54/OR-25–02. Athens, USA: The University of Georgia Center for Education for the International Society for Plant Molecular Biology.
Erickson JM, Rahire M and Rochaix JD (1985) Herbicide resistance and cross-resistance; changes at three distinct sites in the herbicide-binding protein. Science 228: 204–207.
Evans MCW (1987) Plant reaction centre defined. Nature 327: 284–285.
Feher G and Okamura MY (1978) Chemical composition and properties of reaction centers. In: Clayton RK and Sistrom WR (eds) The Photosynthetic Bacteria pp. 349–386. New York: Plenum Press.
Gilbert CW, Williams JGK, Williams KAL and Arntzen CJ (1985) Herbicide action in photosynthetic bacteria. In: Steinbeck KE, Bonitz S, Arntzen CJ and Bogorad L (eds) Molecular Biology of the Photosynthetic Apparatus, pp 67–71. Cold Spring Harbor: Cold Spring Harbor Laboratory.
Golden SS and Haselkorn R (1985) Mutation to herbicide resistance maps within the psbA gene of Anacystis nidulans R2. Science 229: 1104–1107.
Goloubinoff P, Edelman M and Hallick RB (1984) Chloroplast-coded atrazine resistance in Solanum nigrum: psbA loci from susceptible and resistant biotypes are isogenic except for a single codon change. Nucl Acid Res 12: 9489–9496.
Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA Cloning, Vol 1, pp. 109–135. Washington DC: IRL Press.
Hanahan D and Meselson M (1980) Plasmid screening at high colony density. Gene 10, 63–67.
Hirschberg J and McIntosh L (1983) Molecular basis of herbicide resistance in Amaranthus hybridus. Science 222, 1346–1349.
Hirschberg J, Bleecker A, Kyle DJ, McIntosh L and Arntzen CJ (1984) The molecular basis of triazine-herbicide resistance in higher-plant chloroplasts. Z. Naturforsch. 39c, 412–420.
Holmes DS and Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114, 193–197.
Johanningmeier U, Bodner U and Wildner GF (1987) A new mutation in the gene coding for the herbicide-binding protein in Chlamydomonas. FEBS Lett 211, 221–224.
Kleinfeld D, Okamura MY and Feher G (1984) Electron transfer in reaction centers of Rhodopseudomonas sphaeroides. Determination of the charge recombination pathway of D+QAQB - and free energy and kinetic relations between QA -QB and QAQB -. Biochem Biophys Acta 766, 126–140.
Kyle DJ (1985) The 32000 dalton QB protein of photosystem II. Photochem Photobiol 41, 107–116.
Maniatis T, Fritsch EF and Sambrook J (1982) Molecular Cloning-A Laboratory Manual. Cold Spring Harbor: Cold Spring Harbor Press.
McComb JC and Wraight CA (1983) Activity of analogues as primary and secondary quinones in photosynthetic reaction centers (Abstract). Biophys J 41, 39a.
McElroy JD, Mauzerall DC and Feher G (1974) Characterization of primary reactants in bacterial photosynthesis. II. Kinetic studies of the light-induced signal (g=2.0026) and the optical absorbance changes at cryogenic temperatures. Biochem Biophys Acta 333, 261–278.
Messing J (1983) New M13 vectors for cloning. Meth Enz 101, 20–78.
Messing J and Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digested restriction fragments. Gene 19, 269–276.
Michel H, Epp O and Deisenhofer J (1986a) Pigment-protein interactions in the photosynthetic reaction centre from Rhodopseudomonas viridis. EMBO J 5, 2445–2451.
Michel H, Weyer KA, Gruenberg H, Dugner I, Oesterhelt D and Lottspeich F (1986b) The ‘light’ and ‘medium’ subunits of the photosynthetic reaction centre from Rhodopseudomonas viridis: isolation of the genes, nucleotide and amino acid sequence. EMBO J 5, 1149–1158.
Michel-Beyerle ME (ed.) (1985) Antennas and Reaction Centers of Photosynthetic Bacteria: Structure, Interactions, and Dynamics. New York: Springer-Verlag.
Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor: Cold Spring Harbor Press.
Nanba O and Satoh K (1987) Isolation of a photosystem II reaction center consisting of D-1 and D-2 polypetpides and cytochrome b-559. Proc Natl Acad Sci USA 84, 109–112.
Okamura MY (1984) On the herbicide site in bacterial reaction centers. In: Thornber JP, Staehelin LA and Hallick RB (eds) Biosynthesis of the Photosynthetic Apparatus: Molecular Biology, Development and Regulation, pp 381–390. New York: Alan R Liss, Inc.
Okamura MY, Isaacson RA and Feher G (1975) The primary acceptor in bacterial photosynthesis: the obligatory role of ubiquinone in photoactive reaction centers of Rhodopseudomonas sphaeroides. Proc Natl Acad Sci USA 72, 3491–3495.
Okamura MY, Debus RJ, Kleinfeld D and Feher G (1982) Quinone binding sites in reaction centers from photosynthetic bacteria. In: Trumpower BC (ed) Functions of Quinones in Energy Conserving Systems, pp 299–317. New York: Academic Press.
Okamura MY, Abresch EC and Debus RJ (1985) Reaction centers from triazine resistant strains of Rhodopseudomonas sphaeroides: localization of the mutation site by protein hybridization experiments. Biochim Biophys Acta 810, 110–113.
Okamura MY, Satoh K, Isaacson RA and Feher G (1987) Evidence of the primary charge separation in the D1D2 complex of photosystem II from spinach: EPR of the triplet state. In: Biggins J (ed) Progress in Photosynthesis Research, Vol 1, pp I.4.379-I.4.381. Dordrecht: Martinus Nijhoff Publishers.
Ort DR and Melandri BA (1982) Mechanism of ATP synthesis. In: Govindjee (ed.) Photosynthesis: Energy Conversion by Plant and Bacteria, Vol 1, pp 537–587. New York: Academic Press.
Ort DR (1986) Energy transduction in oxygenic photosynthesis: an overview of structure and mechanism. In: Staehelin LA and Arntzen CJ (eds) Encyclopedia of Plant Physiology, Vol 19: Photosynthesis III: Photosynthetic Membranes and Light Harvesting Systems, pp 143–196. New York: Springer-Verlag.
Paddock ML, Williams JC, Rongey SH, Abresch EC, Feher G and Okamura MY (1987) Characterization of three herbicide resistant mutants of Rhodopseudomonas sphaeroides 2.4.1: structure-function relationship. In: Biggins J (ed.) Progress in Photosynthesis Research, Vol 3 pp III.11.775-III.11.778. Dordrecht: Martinus Nijhoff Publishers.
Pfister K and Arntzen CJ (1979) The mode of action of photosystem II-specific inhibitors in herbicide-resistant weed biotypes. Z Naturforsch 34c, 996–1009.
Rawn JD (1983) Biochemistry. New York: Harper and Row Publishers.
Robinson H, Golden S, Brusslan J and Haselkorn R (1987) Functioning of photosystem II in mutant strains of the cyanobacterium Anacystis nidulans R2. In: Biggins J (ed.) Progress in Photosynthesis Research, Vol 4, pp IV.12.825-IV.12.828. Dordrecht: Martinus Nijhoff Publishers.
Sanger F, Nicklen S and Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74, 5463–5467.
Schenck CC, Sistrom WR and Capaldi RA (1986) structure-function studies in reaction centers: characterization of an herbicide resistance mutation in Rhodopseudomonas sphaeroides (Abstract). Biophys J 49, 486a.
Sinning I and Michel H (1987) Sequence analysis of mutants from Rhodopseudomonas viridis resistant to the herbicide terbutryn. Z Naturforsch 42c, 751–754.
Stein RR, Castellvi AL, Bogacz JP and Wraight CA (1984) Herbicide-quinone competition in the acceptor complex of photosynthetic reaction centers from Rhodopseudomonas sphaeroides: a bacterial model for PS-II-herbicide activity in plants. J Cell Biochem 24, 243–259.
Tischer W and Strotmann H (1977) Relationship between inhibitor binding by chloroplasts and inhibition of photosynthetic electron transport. Biochim Biophys Acta 460, 113–125.
Trebst A (1986) The topology of the plastoquinone and herbicide binding peptides of photosystem II in the thylakoid membrane. Z Naturforsch 41c, 240–245.
Trebst A (1987) The three-dimensional structure of the herbicide binding niche on the reaction center polypeptides from photosystem II. Z Naturforsch 42c, 742–750.
Velthuys BR (1981) Electron-dependent competition between platoquinone and inhibitors for binding to photosystem II. FEBS Lett 126, 277–281.
Vermass WFJ, Arntzen CJ, Gu L-Q and Yu C-A (1983) Interactions of herbicide and azidoquinones at a photosystem II binding site in the thylakoid membrane. Biochim Biophys Acta 723, 266–275.
Vermaas WFJ, Renger G and Arntzen CJ (1984) Herbicide/quinone binding interactions in photosystem II. Z Naturforsch 39c, 368–373.
Warncke K, Gunner MR, Braun BS, Yu C-A and Dutton PL (1987) Effects of hydrocarbon tail structure on the affinity of substituted quinones for the QA and QB sites in reaction center protein of Rhodopseudomonas sphaeroides R26 (Abstract). Biophys J 51, 124a..
Williams JC, Steiner LA, Ogden RC, Simon MI and Feher G (1983) Primary structure of the M subunit of the reaction center from Rhodopseudomonas sphaeroides. Proc Natl Acad Sci USA 80, 6505–6509.
Williams JC, Steiner LA, Feher G and Simon MI (1984) Primary structure of the L subunit of the reaction center from Rhodopseudomonas sphaeroides. Proc Natl Acad Sci USA 81, 7303–7307.
Williams JC, Steiner LA and Feher G (1986) Primary structure of the reaction center from Rhodopseudomonas sphaeroides. Proteins 1, 312–325.
Wraight CA (1981) Oxidation-reduction physical chemistry of the acceptor quinone complex in bacterial photosynthetic reaction centers: evidence for a new model of herbicide activity. Israel J Chem 21, 348–354.
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.
Youvan DC, Bylina EJ, Alberti M, Begusch H, and Hearst JE (1984) Nucleotide and deduced polypeptide sequences of the photosynthetic reaction center, B870 antenna, and flanking polypeptides from Rhodopseudomonas capsulata. Cell 37, 949–957.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Paddock, M.L., Rongey, S.H., Abresch, E.C. et al. Reaction centers from three herbicide-resistant mutants of Rhodobacter sphaeroides 2.4.1: sequence analysis and preliminary characterization. Photosynth Res 17, 75–96 (1988). https://doi.org/10.1007/BF00047682
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00047682