Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1981 Jun 11;9(11):2563–2576. doi: 10.1093/nar/9.11.2563

Sequence arrangement of the rRNA genes of the dipteran Sarcophaga bullata.

C K French, D L Fouts, J E Manning
PMCID: PMC326872  PMID: 6269054

Abstract

Velocity sedimentation studies of RNA of Sarcophaga bullata show that the major rRNA species have sedimentation values of 26S and 18S. Analysis of the rRNA under denaturing conditions indicates that there is a hidden break centrally located in the 26S rRNA species. Saturation hybridization studies using total genomic DNA and rRNA show that 0.08% of the nuclear DNA is occupied by rRNA coding sequences and that the average repetition frequency of these coding sequences is approximately 144. The arrangement of the rRNA genes and their spacer sequences on long strands of purified rDNA was determined by the examination of the structure of rRNa:DNA hybrids in the electron microscope. Long DNA strands contain several gene sets (18S + 26S) with one repeat unit containing the following sequences in order given: (a) An 18S gene of length 2.12 kb, (b) an internal transcribed spacer of length 2.01 kb, which contains a short sequence that may code for a 5.8S rRNA, (c) A 26S gene of length 4.06 kb which, in 20% of the cases, contains an intron with an average length of 5.62 kb, and (d) an external spacer of average length of 9.23 kb.

Full text

PDF
2563

Images in this article

Selected References

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

  1. Arnheim N., Southern E. M. Heterogeneity of the ribosomal genes in mice and men. Cell. 1977 Jun;11(2):363–370. doi: 10.1016/0092-8674(77)90053-8. [DOI] [PubMed] [Google Scholar]
  2. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  3. Barnett T., Rae P. M. A 9.6 kb intervening sequence in D. virilis rDNA, and sequence homology in rDNA interruptions of diverse species of Drosophila and other diptera. Cell. 1979 Apr;16(4):763–775. doi: 10.1016/0092-8674(79)90092-8. [DOI] [PubMed] [Google Scholar]
  4. Beckingham K., White R. The ribosomal DNA of Calliphora erythrocephala; and analysis of hybrid plasmids containing ribosomal DNA. J Mol Biol. 1980 Mar 15;137(4):349–373. doi: 10.1016/0022-2836(80)90162-x. [DOI] [PubMed] [Google Scholar]
  5. Birnstiel M. L., Wallace H., Sirlin J. L., Fischberg M. Localization of the ribosomal DNA complements in the nucleolar organizer region of Xenopus laevis. Natl Cancer Inst Monogr. 1966 Dec;23:431–447. [PubMed] [Google Scholar]
  6. Borst P., Grivell L. A. The mitochondrial genome of yeast. Cell. 1978 Nov;15(3):705–723. doi: 10.1016/0092-8674(78)90257-x. [DOI] [PubMed] [Google Scholar]
  7. Brown D. D., Weber C. S. Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4 s RNA, 5 s RNA and ribosomal RNA. J Mol Biol. 1968 Jun 28;34(3):661–680. doi: 10.1016/0022-2836(68)90188-5. [DOI] [PubMed] [Google Scholar]
  8. Cech T. R., Rio D. C. Localization of transcribed regions on extrachromosomal ribosomal RNA genes of Tetrahymena thermophila by R-loop mapping. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5051–5055. doi: 10.1073/pnas.76.10.5051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Commerford S. L. Iodination of nucleic acids in vitro. Biochemistry. 1971 May 25;10(11):1993–2000. doi: 10.1021/bi00787a005. [DOI] [PubMed] [Google Scholar]
  10. Faye G., Dennebouy N., Kujawa C., Jacq C. Inserted sequence in the mitochondrial 23S ribosomal RNA gene of the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1979 Jan 5;168(1):101–109. doi: 10.1007/BF00267939. [DOI] [PubMed] [Google Scholar]
  11. Gerbi S. A. Localization and characterization of the ribosomal RNA cistrons in Sciara coprophila. J Mol Biol. 1971 Jun 14;58(2):499–511. doi: 10.1016/0022-2836(71)90367-6. [DOI] [PubMed] [Google Scholar]
  12. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  13. Glover D. M., Hogness D. S. A novel arrangement of the 18S and 28S sequences in a repeating unit of Drosophila melanogaster rDNA. Cell. 1977 Feb;10(2):167–176. doi: 10.1016/0092-8674(77)90212-4. [DOI] [PubMed] [Google Scholar]
  14. Greenberg J. R. Synthesis and properties of ribosomal RNA in Drosophila. J Mol Biol. 1969 Nov 28;46(1):85–98. doi: 10.1016/0022-2836(69)90059-x. [DOI] [PubMed] [Google Scholar]
  15. Heckman J. E., RajBhandary U. L. Organization of tRNA and rRNA genes in N. crassa mitochondria: intervening sequence in the large rRNA gene and strand distribution of the RNA genes. Cell. 1979 Jul;17(3):583–595. doi: 10.1016/0092-8674(79)90266-6. [DOI] [PubMed] [Google Scholar]
  16. Jackson A. O., Larkins B. A. Influence of Ionic Strength, pH, and Chelation of Divalent Metals on Isolation of Polyribosomes from Tobacco Leaves. Plant Physiol. 1976 Jan;57(1):5–10. doi: 10.1104/pp.57.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jolly D. J., Thomas C. A., Jr Nuclear RNA transcripts from Drosophila melanogaster ribosomal RNA genes containing introns. Nucleic Acids Res. 1980 Jan 11;8(1):67–84. doi: 10.1093/nar/8.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jordan B. R., Jourdan R., Jacq B. Late steps in the maturation of Drosophila 26 S ribosomal RNA: generation of 5-8 S and 2 S RNAs by cleavages occurring in the cytoplasm. J Mol Biol. 1976 Feb 15;101(1):85–105. doi: 10.1016/0022-2836(76)90067-x. [DOI] [PubMed] [Google Scholar]
  19. Karrer K. M., Gall J. G. The macronuclear ribosomal DNA of Tetrahymena pyriformis is a palindrome. J Mol Biol. 1976 Jun 25;104(2):421–453. doi: 10.1016/0022-2836(76)90280-1. [DOI] [PubMed] [Google Scholar]
  20. Lava-Sanchez P. A., Puppo S. Occurrence in vivo of "hidden breaks" at specific sites of 26 S ribosomal RNA of Musca carnaria. J Mol Biol. 1975 Jun 15;95(1):9–20. doi: 10.1016/0022-2836(75)90331-9. [DOI] [PubMed] [Google Scholar]
  21. Leon W., Fouts D. L., Manning J. Sequence arrangement of the 16S and 26S rRNA genes in the pathogenic haemoflagellate Leishmania donovani. Nucleic Acids Res. 1978 Feb;5(2):491–504. doi: 10.1093/nar/5.2.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Loening U. E. Molecular weights of ribosomal RNA in relation to evolution. J Mol Biol. 1968 Dec;38(3):355–365. doi: 10.1016/0022-2836(68)90391-4. [DOI] [PubMed] [Google Scholar]
  23. Long E. O., Dawid I. B. Expression of ribosomal DNA insertions in Drosophila melanogaster. Cell. 1979 Dec;18(4):1185–1196. doi: 10.1016/0092-8674(79)90231-9. [DOI] [PubMed] [Google Scholar]
  24. Maizels N. Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976 Nov;9(3):431–438. doi: 10.1016/0092-8674(76)90088-x. [DOI] [PubMed] [Google Scholar]
  25. Manning J. E., Hershey N. D., Broker T. R., Pellegrini M., Mitchell H. K., Davidson N. A new method of in situ hybridization. Chromosoma. 1975 Nov 24;53(2):107–117. doi: 10.1007/BF00333039. [DOI] [PubMed] [Google Scholar]
  26. Manning J., Pellegrini M., Davidson N. A method for gene enrichment based on the avidin-biotin interaction. Application to the Drosophila ribosomal RNA genes. Biochemistry. 1977 Apr 5;16(7):1364–1370. doi: 10.1021/bi00626a020. [DOI] [PubMed] [Google Scholar]
  27. Manning R. F., Samols D. R., Gage L. P. The genes for 18S, 5.8S and 28S ribosomal RNA of Bombyx mori are organized into tandem repeats of uniform length. Gene. 1978 Oct;4(2):153–166. doi: 10.1016/0378-1119(78)90027-6. [DOI] [PubMed] [Google Scholar]
  28. Molgaard H. V., Matthews H. R., Bradbury E. M. Organisation of genes for ribosomal RNA in Physarum polycephalum. Eur J Biochem. 1976 Sep 15;68(2):541–549. doi: 10.1111/j.1432-1033.1976.tb10842.x. [DOI] [PubMed] [Google Scholar]
  29. Pellegrini M., Holmes D. S., Manning J. Application of the avidin-biotin method of gene enrichment to the isolation of long double-stranded DNA containing specific gene sequences. Nucleic Acids Res. 1977 Sep;4(9):2961–2973. doi: 10.1093/nar/4.9.2961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pellegrini M., Manning J., Davidson N. Sequence arrangement of the rDNA of Drosophila melanogaster. Cell. 1977 Feb;10(2):213–214. doi: 10.1016/0092-8674(77)90215-x. [DOI] [PubMed] [Google Scholar]
  31. Renkawitz R., Gerbi S. A., Glätzer K. H. Ribosomal DNA of fly Sciara coprophila has a very small and homogeneous repeat unit. Mol Gen Genet. 1979 May 23;173(1):1–13. doi: 10.1007/BF00267685. [DOI] [PubMed] [Google Scholar]
  32. Samols D., Swift H. Genomic organization in the flesh fly Sarcophaga bullata. Chromosoma. 1979 Nov;75(2):129–143. doi: 10.1007/BF00292203. [DOI] [PubMed] [Google Scholar]
  33. Speirs J., Birnstiel M. Arrangement of the 5-8 S RNA cistrons in the genome of Xenopus laevis. J Mol Biol. 1974 Aug 5;87(2):237–256. doi: 10.1016/0022-2836(74)90146-6. [DOI] [PubMed] [Google Scholar]
  34. Tartof K. D. Evolution of transcribed and spacer sequences in the ribosomal RNA genes of Drosophila. Cell. 1979 Jul;17(3):607–614. doi: 10.1016/0092-8674(79)90268-x. [DOI] [PubMed] [Google Scholar]
  35. Wellauer P. K., Dawid I. B., Brown D. D., Reeder R. H. The molecular basis for length heterogeneity in ribosomal DNA from Xenopus laevis. J Mol Biol. 1976 Aug 25;105(4):461–486. doi: 10.1016/0022-2836(76)90229-1. [DOI] [PubMed] [Google Scholar]
  36. Wellauer P. K., Dawid I. B., Tartof K. D. X and Y chromosomal ribosomal DNA of Drosophila: comparison of spacers and insertions. Cell. 1978 Jun;14(2):269–278. doi: 10.1016/0092-8674(78)90113-7. [DOI] [PubMed] [Google Scholar]
  37. Wellauer P. K., Dawid I. B. The structural organization of ribosomal DNA in Drosophila melanogaster. Cell. 1977 Feb;10(2):193–212. doi: 10.1016/0092-8674(77)90214-8. [DOI] [PubMed] [Google Scholar]
  38. White R. L., Hogness D. S. R loop mapping of the 18S and 28S sequences in the long and short repeating units of Drosophila melanogaster rDNA. Cell. 1977 Feb;10(2):177–192. doi: 10.1016/0092-8674(77)90213-6. [DOI] [PubMed] [Google Scholar]
  39. Wild M. A., Gall J. G. An intervening sequence in the gene coding for 25S ribosomal RNA of Tetrahymena pigmentosa. Cell. 1979 Mar;16(3):565–573. doi: 10.1016/0092-8674(79)90030-8. [DOI] [PubMed] [Google Scholar]
  40. Wu M., Davidson N. Use of gene 32 protein staining of single-strand polynucleotides for gene mapping by electron microscopy: application to the phi80d3ilvsu+7 system. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4506–4510. doi: 10.1073/pnas.72.11.4506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zimmerman J. L., Fouts D. L., Manning J. E. Evidence for a complex class of nonadenylated mRNA in Drosophila. Genetics. 1980 Jul;95(3):673–691. doi: 10.1093/genetics/95.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES