Abstract
We report here the isolation and nucleotide sequence of a cDNA clone encoding a phtosystem I polypeptide that is recognized by a polyclonal antibody prepared against subunit II of the photosystem I reaction center. The transit peptide processing site was determined to occur after Met50 by N terminal sequencing. The decuced sequence of this protein predicts that the polypeptide has a net positive charge (pI=9.6) and no membrane spanning regions are evident from the hydropathy plot. Based on these considerations and the fact that subunit II is solubilized by alkali treatment of thylakoids, we concluded that subunit II is an extrinsic membrane protein. The absence of hydrophobic regions characteristic of thylakoid transfer domains furthermore implies that subunit II is localized on the stromal side of the membrane.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Aebersold RH, Teplow DB, Hood LE, Kent SB: Electroblotting onto activated glass. High efficiency preparation of proteins from analytical sodium dodecyl sulfate acrylamide gels for direct sequence analysis. J Biol Chem 261: 4229–4238 (1986).
Akerlund HE, Jansson C: Localization of a 34 000 and a 23 000 M r polypeptide to the lumenal side of the thylakoid membrane. FEBS Lett 124: 229–232 (1981).
Alexander DC, McKnight TD, Williams BG: A simplified and efficient vector-primer cDNA cloning system. Gene 31: 79–89 (1984).
Bengis C, Nelson N: Purification and properties of the photosystem I reaction center from chloroplasts. J Biol Chem 250: 2783–2788 (1975).
Bengis C, Nelson N: Subunit structure of chloroplast photosystem I reaction center. J Biol Chem 252: 4564–4569 (1977).
Bonnerjea J, Ortiz W, Malkin R: Identification of a 19 kDa polypeptide as an Fe-S center apoprotein in the photosystem I primary electron acceptor complex. Arch Biochem Biophys 240: 15–20 (1985).
Cashmore AR: The isolation of poly A + messenger RNA from higher plants. In: Edelman M, Hallick RB, Chua NH (eds) Methods in Chloroplast Molecular Biology. Elsevier Biomedical Press, Amsterdam (1982) pp. 387–392.
Darr SC, Sommerville SC, Arntzen CJ: Monoclonal antibodies to the light harvesting chlorophyll a/b protein complex of photosystem II. J Cell Biol 103: 733–740 (1986).
DellaPenna D, Alexander DC, Bennett AB: Molecular cloning of tomato fruit polygalacturonase: analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci 83: 6420–6424 (1986).
Fish LE, Kuck U, Bogorad L: Two partially homologous adjacent light inducible maize chloroplast genes encoding polypeptides of the P700 chlorophyll a-protein complex of photosystem I. J Biol Chem 3: 1413–1421 (1985).
Golbeck JH, Cornelius JM: Photosystem I charge separation in the absence of centers A and B. I. Optical characterization of center “A2” and evidence for its association with a 64 kDa peptide. Biochim Biophys Acta 849: 16–24 (1986).
Hayashida N, Matsubayashi T, Shinozaki K, Sugiura M, Inoue K, Hiyama T: The gene for the 9 kD polypeptide, a possible apoprotein for the iron-sulfur centers A and B of the photosystem I complex, in tobacco chloroplast DNA. Curr Genet 12: 247–250 (1987).
Hoffman NE, Pichersky E, Malik VS, Castresana C, Ko K, Darr SC, Cashmore AR: A cDNA clone encoding a photosystem I protein with homology to photosystem II chlorophyll a/b-binding polypeptide. Proc Natl Acad Sci (in press)
Høj PB, Halkier BA, Møller BL: Analysis of isolated PS I polypeptides for acid labile sulfide. In: Biggins J (ed.) Progress in Photosynthesis Research. Martinus Nijhoff, Dordrecht (1987) Vol II. pp. 53–56.
Høj PB, Møller BL: The 110 kDa reaction center protein of photosystem I, P700-chlorophyll a-protein 1, is an iron-sulfur protein. J Biol Chem 261: 14292–14300.
Kirsch W, Seyer P, Herrmann RG: Nucleotide sequence of the clustered genes for two P700 chlorophyll a apoproteins of the photosystem I reaction center and the ribosomal protein S14 of the spinach plastid chromosome. Curr Genet 10: 843–855 (1986).
Kyte J, Doolittle RF: A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105–132 (1982).
Lagoutte B, Setif P, Duranton J: Tentative identification of the apoproteins of iron-sulfur centers of photosystem I. FEBS Lett 174: 24–29 (1984).
Lehmbeck J, Rasmussen OF, Bookjans GB, Jepsen BR, Stummann BM, Henningsen KW: Sequence of two genes in pea chloroplast DNA coding for 84 and 82 kD polypeptides of the photosystem I complex. Plant Mol Biol 7: 3–10 (1986).
Malkin R: Photosystem I. Ann Rev Plant Physiol 33: 455–479 (1982).
Maniatis T, Fritsch EF, Sambrook J: In: tMolecular Cloning — a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., pp. 200–201.
Møller BL, Halkier BA, Høj PB: The organization of the Fe-S acceptors of photosystem I. In: Biggins J (ed.) Progress in Photosynthesis Research. Martinus Nijhoff, Dordrecht (1987) Vol. II, pp. 49–52.
Mullet JE, Burke JJ, Arntzen CJ: Chlorophyll proteins of photosystem I. Plant Physiol 65: 814–822 (1980).
Nechushtai R, Muster P, Binder A, Liveanu V, Nelson N: Photosystem I reaction center from the thermophilic cyanobacterium Mastigocladus laminosus. Proc Natl Acad Sci 80: 1179–1183 (1983).
Nechushtai R, Nelson N: Purification, properties, and biogenesis of Chlamydomonas reinhardii photosystem I reaction center. J Biol Chem 256: 11624–11628 (1981).
Nechushtai R, Nelson N, Mattoo AK, Edelman M: Site of synthesis of subunits to photosystem I reaction center and the proton-ATPase in Spirodela. FEBS Lett 125: 115–119 (1981).
Oh-oka H, Takahashi Y, Wada K, Matsubara H, Ohyama K, Ozeki H: The 8 kDa polypeptide in photosystem I is a probable candidate of an iron-sulfur center protein coded by the chloroplast gene frxA. FEBS Lett 218: 52–54 (1987).
Ohyama K, Fukuzawa H, Kohchi T, Shirai H, Sano T, Sano S, Umesono K, Shiki Y, Takeuchi M, Chang Z, Aota S, Inokuchi H, Ozeki H: Chloroplast organization deduced from complete sequence of liverwort Marchantia polymorpha chloroplast DNA. Nature 322: 572–574 (1986).
Ortiz W, Lam E, Chollar S, Munt D, Malkin R: Topography of the protein complexes of the chloroplast thylakoid membrane. Studies of photosystem I using a chemical probe and proteolytic digestion. Plant Physiol 77: 389–397 (1985).
Piccioni R, Bellemare G, Chua NH: Methods of polyacrylamide gel electrophoresis in the analysis and preparation of plant polypeptides. In: Edelman M, Hallick RB, Chua NH (eds) Methods in Chloroplast Biology. Elsevier Biomedical Press, Amsterdam (1982) pp. 985–1014.
Pichersky E, Hoffman NE, Malik VS, Bernatzky R, Tanksley SD, Szabo L, Cashmore AR: The tomato Cab-4 and Cab-5 genes encode a second type of CAB polypeptides localized in photosystem II. Plant Mol Biol 9: 109–120 (1987).
Rother C, Jansen T, Tyagi A, Tittgen J, Herrmann RG: Plastocyanin is encoded by an uninterrupted nuclear gene in spinach. Curr Genet 11: 171–176 (1986).
Sakurai H, San Pietro A: Association of Fe-S center(s) with the large subunit(s) of photosystem I particles. J Biochem 98: 69–76 (1985).
Schmidt GW, Bartlett SG, Grossman AR, Cashmore AR, Chua NH: Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol 91: 468–478 (1981).
Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse J, Obokata J, Yamaguchi-Shinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M: The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J 5: 2043–2049 (1986).
Smeekens S, de Groot M, van Binsbergen J, Weisbeek P: Sequence of the precursor of the chloroplast thylakoid lumen protein plastocyanin. Nature 317: 456–458 (1985).
Sutton A, Sieburth LE, Bennett J: Light-dependent accumulation and localization of photosystem II proteins in maize. Eur J Biochem 164: 571–578 (1987).
Tittgen J, Hermans J, Steppuhn J, Jansen T, Jansson C, Andersson B, Nechushtai R, Nelson N, Herrmann RG: Isolation of cDNA clones for fourteen nuclear encoded thylakoid membrane proteins. Mol Gen Genet 204: 258–265 (1986).
Tyagi A, Hermans J, Steppuhn J, Jansson C, Vater F, Herrmann RG: Nucleotide sequence of cDNA clones encoding the complete “33 kDa” precursor protein associated with photosynthetic oxygen-evolving complex from spinach. Mol Gen Genet 207: 288–293 (1987).
Westhoff P, Alt J, Nelson N, Bottomley W, Bunemann H, Herrmann RG: Genes and transcripts for the P700 chlorophyll a apoprotein and subunit 2 of the photosystem I reaction center complex from spinach thylakoid membranes. Plant Mol Biol 2: 95–107 (1983).
Yuen S, Hunkapiller MW, Wilson KJ, Yuan PM: SDS-PAGE Electroblotting. Applied Biosystems User Bulletin 25 (1986).
Zanetti G, Merati G: Interaction between photosystem I and ferredoxin. Identification by chemical cross-linking of the polypeptide which binds ferredoxin. Eur J Biochem 169: 143–146 (1987).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hoffman, N.E., Pichersky, E., Malik, V.S. et al. Isolation and sequence of a tomato cDNA clone encoding subunit II of the photosystem I reaction center. Plant Mol Biol 10, 435–445 (1988). https://doi.org/10.1007/BF00014949
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00014949