Summary
Experiments were undertaken to characterize the cytoplasmic ribosomal proteins (r-proteins) in Chlamydomonas reinhardtii and to compare immunologically several cytoplasmic r-proteins with those of chloroplast ribosomes of this alga, Escherichia coli, and yeast. The large and small subunits of the C. reinhardtii cytoplasmic ribosomes were shown to contain, respectively, 48 and 45 r-proteins, with apparent molecular weights of 12,000–59,000. No cross-reactivity was seen between antisera made against cytoplasmic r-proteins of Chlamydomonas and chloroplast r-proteins, except in one case where an antiserum made against a large subunit r-protein cross-reacted with an r-protein of the small subunit of the chloroplast ribosome. Antisera made against one out of five small subunit r-proteins and three large subunit r-proteins recognized r-proteins from the yeast large subunit. Each of the yeast r-proteins has been previously identified as an rRNA binding protein. The antiserum to one large subunit r-protein cross-reacted with specific large subunit r-proteins from yeast and E. coli.
Similar content being viewed by others
References
Bartlett SG, Harris EH, Grabowy CT, Gillham NW, Boynton JE (1979) Ribosomal subunits affected by antibiotic resistance mutations at seven chloroplast loci in Chlamydomonas reinhardtii. Mol Gen Genet 176:199–208
Bartsch M (1985) Correlation of chloroplast and bacterial ribosomal proteins by cross-reactions of antibodies specific to purified Escherichia coli ribosomal proteins. J Biol Chem 260:237–241
Bartsch M, Kimura M, Subramanian AR (1982) Purification, primary structure, and homology relationships of a chloroplast ribosomal protein. Proc Natl Acad Sci USA 79:6871–6875
Bourque DP, Boynton JE, Gillham NW (1971) Studies on the structure and cellular location of various ribosome and ribosomal RNA species in the green alga Chlamydomonas reinhardtii. J Cell Sci 8:153–183
Boynton JE, Gillham NW, Lambowitz AM (1980) Biogenesis of chloroplast and mitochondrial ribosomes. In: Chambliss G, Craven GR, Davies J, Davis K, Kahan L, Nomura M (eds) Ribosomes: structure, function, and genetics. University Park Press, Baltimore, pp 903–950
Brugger M, Boschetti A (1975) Two-dimensional gel electrophoresis of ribosomal proteins from streptomycin-sensitive and streptomycin-resistant mutants of Chlamydomonas reinhardtii. Eur J Biochem 58:603–610
Burnette WN (1981) “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203
Capel MS, Bourque DP (1982) Characterization of Nicotiana tabacum chloroplast and cytoplasmic ribosomal proteins. J Biol Chem 257:7746–7755
Chua NH, Blobel G, Siekevitz P (1973), Characterization of the ribosomal proteins from Schizosaccharomyces pombe by two-dimensional polyacrylamide gel electrophoresis: demonstration that a cycloheximide resistant strain, cyhl, has an altered 60S ribosomal protein. Mol Gen Genet 158:93–100
Conde MF, Boynton JE, Gillham NW, Harris EH, Tingle CL, Wang WL (1975) Chloroplast genes in Chlamydomonas affecting organelle ribosomes: genetic and biochemical analysis of antibiotic-resistant mutants at several gene loci. Mol Gen Genet 140:183–220
Curtis SE, Clegg MT (1984) Molecular evolution of chloroplast DNA sequences. Mol Biol Evol 1:291–301
Dorne AM, Eneas-Filho J, Heizmann P, Mache R (1984) Comparison of ribosomal proteins of chloroplast from spinach and of E. coli. Mol Gen Genet 193:129–134
El-Baradi TTAL, Raue HA, DeRegt VCHR, Planta RJ (1984) Stepwise dissociation of yeast 60S ribosomal subunits by LiCl and identification of L25 as a primary 26S rRNA binding protein. Eur J Biochem 144:393–400
El-Baradi TTAL, Raue HA, Linnekamp M, Planta RJ (1985) Identification of yeast 60S ribosomal proteins cross-linked to rRNA by 2-iminothiolane. FEBS Lett 186:26–30
Eneas-Filho J, Hartley MR, Mache R (1981) Pea chloroplast ribosomal proteins: characterization and site of synthesis. Mol Gen Genet 184:484–488
Fischer N, Stoffler G, Wool IG (1978) Immunological comparisons of the proteins of chicken and rat liver ribosomes. J Biol Chem 253:7355–7360
Freyssinet G (1977) The protein synthesizing system of Euglena: synthesis of ribosomal proteins in vivo and their characterization. Physiol Veg 15:519–550
Gillham NW (1978) Organelle heredity. Raven Press, New York, pp 95–96
Gualerzi C, Janda HG, Passow H, Stoffler G (1974) Studies on the protein moiety of plant ribosomes. J Biol Chem 249:3347–3355
Hancock K, Tsang VCW (1983) India ink staining of proteins on nitrocellulose paper. Anal Biochem 133:157–162
Hanson MR, Davidson JN, Mets LJ, Bogorad L (1974) Characterization of chloroplast and cytoplasmic ribosomal proteins of Chlamydomonas reinhardtii by two-dimensional gel electrophoresis. Mol Gen Genet 132:105–118
Hoober JK, Blobel G (1969) Characterization, of the chloroplastic and cytoplasmic ribosomes of Chlamydomonas reinhardtii. J Mol Biol 41:121–138
Howard GA, Traugh JA, Croser EA, Traut RR (1975) Ribosomal proteins from rabbit reticulocytes: number and molecular weights of proteins from ribosomal subunits. J Mol Biol 93:391–404
Howard GA, Smith RL, Gordon J (1976) Chicken liver ribosomes: characterization of cross-reaction and inhibition of some functions by antibodies prepared against Escherichia coli ribosomal proteins L7 and L12. J Mol Biol 106:623–637
Kalb VF, Jr, Bernlohr RW (1977) A new spectrophotometric assay for protein in cell extracts. Anal Biochem 82:362–371
Kaltschmidt E, Wittmann HG (1972) Ribosomal proteins XXXII. Comparison of several extraction methods for proteins from Escherichia coli ribosomes. Biochimie 54:167–175
Kozak M (1983) Comparison of initiation of protein synthesis in prokaryotes, eukaryotes, and organelles. Microbiol Rev 47:1–45
Kreutzfeldt C, Neumann T, Dierig A (1986) Immunological homologies between ribosomal proteins amongst lower eukaryotes. Curr Genet 10:537–544
Kruiswijk J, Planta RJ (1974) Analysis of the protein of yeast ribosomal subunits by two-dimensional polyacrylamide gel electrophoresis. Mol Biol Rep 1:409–415
Krystosek A, Bitte LF, Cawthon ML, Kabat D (1974) Phosphorylation of ribosomal proteins in eukaryotes. In: Nomura M, Tissieres A, Lengyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 855–870
Lake JA (1983) Ribosome evolution: the structural bases of protein synthesis in archaebacteria, eubacteria, and eukaryotes. Prog Nucleic Acid Res Mol Biol 30:163–194
Lee JC, Henry B, Yeh YC (1983) Binding of proteins from the large ribosomal subunits to 5.8S rRNA of Saccharomyces cerevisiae. J Biol Chem 258:854–858
LeGoff V, Perrot M, Begueret J (1984) Electrophoretical and immunological comparison of the ribosomal, proteins of the fungus Podospora anserina and the yeast Saccharomyces cerevisiae. Curr Genet 9:53–58
Lennox ES (1955) Transduction of linked genetic characters of the host by bacteriophage P1. Virology 1:190–206
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Matheson AT, Yaguchi M (1981) The ribosome as a phylogenetic probe. In: Schweiger HG (ed) International cell biology 1980–1981. Springer, New York, pp 103–110
Mets LJ, Bogorad L (1974) Two-dimensional polyacrylamide gel electrophoresis: an improved method for ribosomal proteins. Anal Biochem 57:200–210
Michel S, Traut RR, Lee JC (1983) Yeast ribosomal proteins: electrophoretic analysis in four two-dimensional gel systems-correlation of nomenclatures. Mol Gen Genet 191:251–256
Moldave K (1985) Eukaryotic protein synthesis. Annu Rev Biochem 54:1109–1149
Moller W, Slobin LI, Amons R, Richter D (1975) Isolation and characterization of two acidic proteins of 60S ribosomes from Artemia salina cysts. Proc Natl Acad Sci USA 72:4744–4748
Nazar RN, Yaguchi M, Willick GE (1982) The 5S RNA-protein complex from yeast: a model for the evolution and structure of the eukaryotic ribosome. Can J Biochem 60:490–496
Nierhaus KH (1980) Analysis of the assembly and function of the 50S subunit from Escherichia coli ribosomes by reconstitution. In: Chambliss G, Craven GR, Davies J, Davis K, Kahan L, Nomura M (eds) Ribosomes: structure, function, and genetics. University Park Press, Baltimore, pp 267–294
Nierhaus KH, Dohme F (1979) Total reconstitution of 50S subunits from Escherichia coli ribosomes. Methods Enzymol 59:443–449
Otaka E, Kobata K (1978) Yeast ribosomal proteins: I. Characterization of cytoplasmic ribosomal proteins by two-dimensional gel electrophoresis. Mol Gen Genet 162:259–268
Otaka E, Osawa S (1981) Yeast ribosomal proteins: V. Correlation of several nomenclatures and proposal of a standard nomenclature. Mol Gen Genet 181:176–182
Otaka E, Higo K, Itoh T (1983) Yeast ribosomal proteins: VII. Cytoplasmic ribosomal proteins from Schizosaccharomyces pombe. Mol Gen Genet 191:519–524
Reyes R, Vazquez D, Ballesta JPG (1978) Structure of the yeast ribosomes: proteins associated with the rRNA. Biochim Biophys Acta 521:229–234
Schmid G, Strobel O, Stoffler-Meilicke M, Stoffler G, Bock A (1984) A ribosomal protein that is immunologically conserved in archaebacteria, eubacteria, and eukaryotes. FEBS Lett 177:189–194
Schmidt RJ, Richardson CB, Gillham NW, Boynton JE (1983) Sites of synthesis of chloroplast ribosomal proteins in Chlamydomonas. J Cell Biol 96:1451–1463
Schmidt RJ, Myers AM, Gillham NW, Boynton JE (1984) Immunological similarities between specific chloroplast ribosomal proteins from Chlamydomonas reinhardtii and ribosomal proteins from Escherichia coli. Mol Biol Evol 1:317–334
Schmidt RJ, Hosler JP, Gillham NW, Boynton JE (1985) Biogenesis and evolution of chloroplast ribosomes: cooperation of nuclear and chloroplast genes. In: Steinback K, Bonitz S, Arntzen C, Bogorad L (eds) The molecular biology of the photosynthetic apparatus. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 417–427
Sherman F, Fink GR, Hicks JB (1983) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 61–64
Sikorski MM, Przybyl D, Legocki AB, Kudlicki W, Gasior E, Zajac J, Borkowski T (1979) The ribosomal proteins of wheat germ: distribution and phosporylation, in vitro. Plant Sci Lett 15:387–397
Sikorski MM, Przybyl D, Legocki AB, Nierhaus KH (1983) Group fractionation of wheat germ ribosomal proteins. Plant Sci Lett 30:303–320
Stoffler G, Wool IG, Lin A, Rak K-H (1974) The identification of the eukaryotic ribosomal proteins homologous, with Escherichia coli proteins L7 and L12. Proc Natl Acad Sci USA 71:4723–4726
Sueoka N (1960) Mitotic replication of deoxyribonucleic acid in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 46:83–89
Svoboda AJ, McConkey EH (1981) Identification of homologous ribosomal proteins in HeLa cells and in Tetrahymena pyriformis: a study of proteins binding 5S RNA and acidic proteins released from 40S subunits by EDTA. Eur J Biochem 116:397–402
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Vazquez D, Jimenez A (1980) Antibiotic inhibitors of translation in eukaryotes. In: Chambliss G, Craven GR, Davies J, Davis K, Kahan L, Nomura M (eds) Ribosomes: structure, function, and genetics. University Park Press, Baltimore, pp 847–869
Warner JR (1982) The yeast ribosome: structure, function, and synthesis. In: Strathern JN, Jones EW, Broach JR (eds) Molecular biology of the yeast Saccharomyces: metabolites and gene expression. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 529–560
Wittmann HG (1974) Purification and identification of Escherichia coli ribosomal proteins. In: Nomura M, Tissieres A, Lengyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 93–114
Wittmann HG, Littlechild JA, Wittmann-Liebold B (1979) Structure of ribosomal proteins. In: Chambliss, G, Craven G, Davies J, Davis D, Kahan L, Nomura M (eds) Ribosomes: structure, function, and genetics. University Park Press, Baltimore, pp 51–88
Wool IG (1979) The structure and function of eukaryotic ribosomes. Annu Rev Biochem 48:719–754
Zinker S, Warner JR (1976) The ribosomal proteins of Saccharomyces cerevisiae. Phosphorylated and exchangable protein. J Biol Chem 251:1799–1807
Author information
Authors and Affiliations
Additional information
Communicated by R.G. Herrmann
Rights and permissions
About this article
Cite this article
Fleming, G.H., Boynton, J.E. & Gillham, N.W. Cytoplasmic ribosomal proteins from Chlamydomonas reinhardtii: Characterization and immunological comparisons. Mol Gen Genet 206, 226–237 (1987). https://doi.org/10.1007/BF00333578
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00333578