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. 1990 Aug 25;18(16):4695–4701. doi: 10.1093/nar/18.16.4695

Identification and characterization of a Dictyostelium discoideum ribosomal protein gene.

D E Szymkowski 1, R A Deering 1
PMCID: PMC331922  PMID: 1975664

Abstract

We have identified a developmentally repressed large-subunit ribosomal protein gene of Dictyostelium discoideum based on sequence similarity to other ribosomal proteins. Protein rpl7 is homologous to large subunit ribosomal proteins from the rat and possibly to Mycoplasma capricolum and Escherichia coli, but is not similar to three sequenced ribosomal proteins in Dictyostelium. The rpl7 gene is present at one copy per genome, as are six other cloned Dictyostelium ribosomal proteins. Restriction fragment length polymorphisms exist for ribosomal protein genes rpl7, rp1024, and rp110 in strain HU182; most Dictyostelium ribosomal protein genes examined are linked no closer than 30-100 kb to each other in the genome. Dictyostelium ribosomal proteins are known to be developmentally regulated, and levels of rpl7 transcript gradually decrease during the 24-hour development cycle. This drop correlates with that of rp1024, indicating these and other ribosomal protein genes may be coordinately regulated. To determine the cellular location of the protein, we raised antibodies to an rpl7-derived branched synthetic peptide. These antibodies cross-reacted with one protein of the expected size in a ribosomal protein fraction of Dictyostelium, indicating that the product of gene rpl7 is localized in the ribosome.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Auron P. E., Fahnestock S. R. Functional organization of the large ribosomal subunit of Bacillus stearothermophilus. J Biol Chem. 1981 Oct 10;256(19):10105–10110. [PubMed] [Google Scholar]
  2. Barker W. C., Ketcham L. K., Dayhoff M. O. A comprehensive examination of protein sequences for evidence of internal gene duplication. J Mol Evol. 1978 Feb 21;10(4):265–281. doi: 10.1007/BF01734217. [DOI] [PubMed] [Google Scholar]
  3. Barritault D., Expert-Bezancon A., Guérin M. F., Hayes D. The use of acetone precipitation in the isolation of ribosomal proteins. Eur J Biochem. 1976 Mar 16;63(1):131–135. doi: 10.1111/j.1432-1033.1976.tb10215.x. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Bzik D. J., Li W. B., Horii T., Inselburg J. Molecular cloning and sequence analysis of the Plasmodium falciparum dihydrofolate reductase-thymidylate synthase gene. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8360–8364. doi: 10.1073/pnas.84.23.8360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Deering R. A. Constitutive and gamma ray modified uptake of labelled precursors into the DNA of Dictyostelium discoideum during development. J Gen Microbiol. 1982 Oct;128(10):2439–2447. doi: 10.1099/00221287-128-10-2439. [DOI] [PubMed] [Google Scholar]
  8. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dudov K. P., Perry R. P. The gene family encoding the mouse ribosomal protein L32 contains a uniquely expressed intron-containing gene and an unmutated processed gene. Cell. 1984 Jun;37(2):457–468. doi: 10.1016/0092-8674(84)90376-3. [DOI] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  11. Firtel R. A., Bonner J. Characterization of the genome of the cellular slime mold Dictyostelium discoideum. J Mol Biol. 1972 May 28;66(3):339–361. doi: 10.1016/0022-2836(72)90419-6. [DOI] [PubMed] [Google Scholar]
  12. Geyer P. K., Meyuhas O., Perry R. P., Johnson L. F. Regulation of ribosomal protein mRNA content and translation in growth-stimulated mouse fibroblasts. Mol Cell Biol. 1982 Jun;2(6):685–693. doi: 10.1128/mcb.2.6.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hardy S. J., Kurland C. G., Voynow P., Mora G. The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochemistry. 1969 Jul;8(7):2897–2905. doi: 10.1021/bi00835a031. [DOI] [PubMed] [Google Scholar]
  14. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  15. Kief D. R., Warner J. R. Hierarchy of elements regulating synthesis of ribosomal proteins in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Nov;1(11):1016–1023. doi: 10.1128/mcb.1.11.1016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lin A., Chan Y. L., Jones R., Wool I. G. The primary structure of rat ribosomal protein S12. The relationship of rat S12 to other ribosomal proteins and a correlation of the amino acid sequences of rat and yeast ribosomal proteins. J Biol Chem. 1987 Oct 15;262(29):14343–14351. [PubMed] [Google Scholar]
  17. Lin A., Chan Y. L., McNally J., Peleg D., Meyuhas O., Wool I. G. The primary structure of rat ribosomal protein L7. The presence near the amino terminus of L7 of five tandem repeats of a sequence of 12 amino acids. J Biol Chem. 1987 Sep 15;262(26):12665–12671. [PubMed] [Google Scholar]
  18. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  19. Marck C. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res. 1988 Mar 11;16(5):1829–1836. doi: 10.1093/nar/16.5.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mehdy M. C., Ratner D., Firtel R. A. Induction and modulation of cell-type-specific gene expression in Dictyostelium. Cell. 1983 Mar;32(3):763–771. doi: 10.1016/0092-8674(83)90062-4. [DOI] [PubMed] [Google Scholar]
  21. Monk R. J., Meyuhas O., Perry R. P. Mammals have multiple genes for individual ribosomal proteins. Cell. 1981 May;24(2):301–306. doi: 10.1016/0092-8674(81)90319-6. [DOI] [PubMed] [Google Scholar]
  22. Muto A., Kawauchi Y., Yamao F., Osawa S. Preferential use of A- and U-rich codons for Mycoplasma capricolum ribosomal proteins S8 and L6. Nucleic Acids Res. 1984 Nov 12;12(21):8209–8217. doi: 10.1093/nar/12.21.8209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nabeshima Y. I., Ogata K. Stimulation of the synthesis of ribosomal proteins in regenerating rat liver with special reference to the increase in the amounts of effective mRNAs for ribosomal proteins. Eur J Biochem. 1980 Jun;107(2):323–329. doi: 10.1111/j.1432-1033.1980.tb06032.x. [DOI] [PubMed] [Google Scholar]
  24. Needleman S. B., Wunsch C. D. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970 Mar;48(3):443–453. doi: 10.1016/0022-2836(70)90057-4. [DOI] [PubMed] [Google Scholar]
  25. Pearson N. J., Fried H. M., Warner J. R. Yeast use translational control to compensate for extra copies of a ribosomal protein gene. Cell. 1982 Jun;29(2):347–355. doi: 10.1016/0092-8674(82)90151-9. [DOI] [PubMed] [Google Scholar]
  26. Ramagopal S. Acidic ribosomal proteins of Dictyostelium discoideum that show electrophoretic similarity to Escherichia coli proteins L7 and L12. Biochem Cell Biol. 1989 Oct;67(10):712–718. doi: 10.1139/o89-106. [DOI] [PubMed] [Google Scholar]
  27. Ramagopal S., Ennis H. L. Regulation of synthesis of cell-specific ribosomal proteins during differentiation of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1981 May;78(5):3083–3087. doi: 10.1073/pnas.78.5.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ramagopal S., Ennis H. L. Studies on ribosomal proteins in the cellular slime mold Dictyostelium discoideum. Resolution, nomenclature and molecular weights of proteins in the 40-S and 60-S ribosomal subunits. Eur J Biochem. 1980 Apr;105(2):245–258. doi: 10.1111/j.1432-1033.1980.tb04495.x. [DOI] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Singleton C. K., Delude R. L., McPherson C. E. Characterization of genes which are deactivated upon the onset of development in Dictyostelium discoideum. Dev Biol. 1987 Feb;119(2):433–441. doi: 10.1016/0012-1606(87)90047-9. [DOI] [PubMed] [Google Scholar]
  31. Singleton C. K., Manning S. S., Ken R. Primary structure and regulation of vegetative specific genes of Dictyostelium discoideum. Nucleic Acids Res. 1989 Dec 11;17(23):9679–9692. doi: 10.1093/nar/17.23.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Singleton C. K. Nucleotide sequence of V1, a ribosomal protein gene from Dictyostelium discoideum. Nucleic Acids Res. 1989 Oct 11;17(19):7989–7989. doi: 10.1093/nar/17.19.7989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Steel L. F., Jacobson A. Post-transcriptional regulation of ribosomal protein gene expression during development in Dictyostelium discoideum. Dev Genet. 1988;9(4-5):421–434. doi: 10.1002/dvg.1020090421. [DOI] [PubMed] [Google Scholar]
  34. Steel L. F., Jacobson A. Ribosomal proteins are encoded by single copy genes in Dictyostelium discoideum. Gene. 1986;41(2-3):165–172. doi: 10.1016/0378-1119(86)90095-8. [DOI] [PubMed] [Google Scholar]
  35. Steel L. F., Jacobson A. Translational control of ribosomal protein synthesis during early Dictyostelium discoideum development. Mol Cell Biol. 1987 Mar;7(3):965–972. doi: 10.1128/mcb.7.3.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Szymkowski D. E., Kelly B., Deering R. A. A Dictyostelium discoideum cDNA coding for a protein with homology to the rat ribosomal protein L7. Nucleic Acids Res. 1989 Jul 11;17(13):5393–5393. doi: 10.1093/nar/17.13.5393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tam J. P. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5409–5413. doi: 10.1073/pnas.85.15.5409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Taylor G. R., Lagosky P. A., Storms R. K., Haynes R. H. Molecular characterization of the cell cycle-regulated thymidylate synthase gene of Saccharomyces cerevisiae. J Biol Chem. 1987 Apr 15;262(11):5298–5307. [PubMed] [Google Scholar]
  39. Tushinski R. J., Warner J. R. Ribosomal proteins are synthesized preferentially in cells commencing growth. J Cell Physiol. 1982 Jul;112(1):128–135. doi: 10.1002/jcp.1041120119. [DOI] [PubMed] [Google Scholar]
  40. Wagner M., Perry R. P. Characterization of the multigene family encoding the mouse S16 ribosomal protein: strategy for distinguishing an expressed gene from its processed pseudogene counterparts by an analysis of total genomic DNA. Mol Cell Biol. 1985 Dec;5(12):3560–3576. doi: 10.1128/mcb.5.12.3560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Warrick H. M. Codon frequency in Dictyostelium discoideum. Methods Cell Biol. 1987;28:497–500. [PubMed] [Google Scholar]
  42. Welker D. L., Hirth K. P., Romans P., Noegel A., Firtel R. A., Williams K. L. The use of restriction fragment length polymorphisms and DNA duplications to study the organization of the actin multigene family in Dictyostelium discoideum. Genetics. 1986 Jan;112(1):27–42. doi: 10.1093/genetics/112.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zurawski G., Zurawski S. M. Structure of the Escherichia coli S10 ribosomal protein operon. Nucleic Acids Res. 1985 Jun 25;13(12):4521–4526. doi: 10.1093/nar/13.12.4521. [DOI] [PMC free article] [PubMed] [Google Scholar]

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