Skip to main content
Log in

Organization of the nuclear ribosomal DNA of Chlamydomonas reinhardii

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Hybridization of cytoplasmic ribosomal RNA (rRNA) to restriction endonuclease digests of nuclear DNA of Chlamydomonas reinhardii reveals two BamHI ribosomal fragments of 2.95 and 2.35×106 d and two SalI ribosomal fragments of 3.8 and 1.5×106 d. The ribosomal DNA (rDNA) units, 5.3×106 d in size, appear to be homogeneous since no hybridization of rDNA to other nuclear DNA fragments can be detected. The two BamHI and SalI ribosomal fragments have been cloned and a restriction map of the ribosomal unit has been established. The location of the 25S, 18S and 5.8S rRNA genes has been determined by hibridizing the rRNAs to digests of the ribosomal fragments and by observing RNA/DNA duplexes in the electron microscope. The data also indicate that the rDNA units are arranged in tandem arrays. The 5S rRNA genes are not closely located to the 25S and 18S rRNA genes since they are not contained within the nuclear rDNA unit. In addition no sequence homology is detectable between the nuclear and chloroplast rDNA units of C. reinhardii.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

rRNA:

ribosomal RNA

rDNA:

ribosomal DNA d, dalton

References

  • Allet, B.: Fragments produced by cleavage of λ deoxyribonucleic acid with the Hemophilus parainfluenzae restriction enzyme HpaII. Biochemistry 12, 3972–3977 (1973)

    Google Scholar 

  • Artavanis-Tsakonas, S., Schedl, P., Tschudi, C., Pirrotta, V., Steward, R., Gehring, W.G.: The 5 S genes of Drosophila melanogaste. Cell 12, 1057–1067 (1977)

    Google Scholar 

  • Bastia, D., Chiang, K.S., Swift, H.: Studies on the ribosomal RNA eistrons of chloroplast and nucleus in Chlamydomonas reinhardii. In Abstracts Am. Soc. Cell Biol. p. 25 (1971)

  • Bedbrook, J.R., Kolodner, R., Bogorad, L.: Zea mays chloroplast ribosomal RNA genes are part of a 22,000 base pair inverted repeat. Cell 11, 739–749 (1977)

    Google Scholar 

  • Bell, G.I., Degennaro, L.J., Gelfand, D.L., Bishop, R.J., Valenzuela, P., Rutter, W.J.: Ribosomal RNA genes of Saccharomyces cerevisiae. 1. Physical map of the repeating unit and location of the regions loding for 5 S, 5.8 S, 18 S and 25 S ribosomal RNAs. J. Biol. Chem. 252, 8118–8125 (1977)

    Google Scholar 

  • Berg, D.E., Davies, J., Allet, B., Rochaix, J.D.: Transposition of R factor genes to bacteriophage λ. Proc. Natl. Acad. Sci. U.S.A. 72, 3628–3632 (1975)

    Google Scholar 

  • Berger, S., Schweiger, H.G.: An apparent lack of non-transcribed spacers in rDNA of a green alga. Mol. Gen. Genet. 139, 269–275 (1975)

    Google Scholar 

  • Bolivar, F., Rodriguez, R.L., Betlach, M.C., Boyer, H.W.: Construction and characterization of new cloning vehicles. 1. Ampicillin-resistant derivatives of the plasmid pMB9. Gene 2, 75–93 (1977)

    Google Scholar 

  • Brown, D.B., Weber, C.S.: Gene linkage by RNA-DNA hybridization. Unique DNA sequences homologous to 4 S RNA, 5 S RNA and ribosomal RNA. J. Mol. Biol. 34, 661–680 (1968)

    Google Scholar 

  • Cohen, S.N., Chang, A.C.Y.: Recircularization and autonomous replication of a sheared R-factor segment in Escherichia coli transformants. Proc. Natl. Acad. Sci. U.S.A. 70, 1293–1297 (1973)

    Google Scholar 

  • Gramer, J.H., Farrelly, F.W., Rownd, R.H.: Restriction endonuclease analysis of ribosomal DNA from Saccharomyces cerevisiae. Mol. Gen. Genet. 148, 233–241 (1976)

    Google Scholar 

  • Deonier, R.C., Othsubo, E., Lee, H.J., Davidson, N.: Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. VII. Mapping the ribosomal RNA genes of plasmid F14. J. Mol. Biol. 89, 619–629 (1974)

    Google Scholar 

  • Frankel, G., Cockburn, A.F., Kindler, K.L., Firtel, R.A.: Organization of the ribosomal RNA genes of Dictyostelium discoideum. Mapping of the transcribed region. J. Mol. Biol. 109, 539–558 (1977)

    Google Scholar 

  • Gillespie, D., Spiegelman, S.: A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. Mol. Biol. 12, 829–842 (1965)

    Google Scholar 

  • Grant, D., Chiang, K.S.: Restriction enzyme analysis of the mitochondrial DNA from Chlamydomonas reinhardii. Biophys. J. 17, 284a (1977)

  • Grünstein, M., Hogness, D.S.: Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. U.S.A. 72, 3961–3965 (1975)

    Google Scholar 

  • Howell, S.H.: The differential synthesis and degradation of ribosomal DNA during the vegetative cell cycle in Chlamydomonas reinhardii. Nature New Biol. 240, 264–267 (1972)

    Google Scholar 

  • Karrer, K.M., Gall, J.G.: The macronuclear ribosomal DNA of Tetrahymena pyriformis is a palindrome. J. Mol. Biol. 104, 421–453 (1976)

    Google Scholar 

  • Katz, L., Kingsburg, D.T., Helinski, D.R.: Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J. Bacteriol. 114, 577–591 (1973)

    Google Scholar 

  • Maden, B.E., Robertson, J.S.: Demonstration of the “5.8 S” ribosomal sequence in HeLa cell ribosomal precursor RNA. J. Mol. Biol. 87, 227–235 (1974)

    Google Scholar 

  • Maniatis, T., Jeffrey, A., van de Sande, H.: Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14, 3787–3794 (1975a)

    Google Scholar 

  • Maniatis, T., Jeffrey, A., Kleid, D.G.: Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. U.S.A. 72, 1184–1188 (1975b)

    Google Scholar 

  • Maxam, A.M., Gilbert, W.: A new method for sequencing DNA. Proc. Natl. Acad. Sci. U.S.A. 74, 560–564 (1977)

    Google Scholar 

  • Miller, M.J., McMahon, D.: Synthesis and maturation of chloroplast and cytoplasmic ribosomal RNA in Chlamydomonas reinhardii. Biochim. Biophys. Acta 366, 35–44 (1974)

    Google Scholar 

  • Pene, J.J., Knight, E., Darnell, J.E.: Characterization of a new low molecular weight RNA in HeLa cell ribosomes. J. Mol. Biol. 33, 609–623 (1968)

    Google Scholar 

  • Pigott, G.H., Carr, N.G.: Homology between nucleic acids of blue-green algae and chloroplasts of Euglena gracilis. Science 175, 1259–1261 (1972)

    Google Scholar 

  • Rochaix, J.D.: Restriction endonuclease map of the chloroplast DNA of Chlamydomonas reinhardii. J. Mol. Biol. 126, 597–617 (1978)

    Google Scholar 

  • Rochaix, J.D.: Restriction fragments from Chlamydomonas chloroplast DNA. In: Methods in enzymology (Grossman, L. and Moldave, K., eds.). New York: Academic Press (1980)

    Google Scholar 

  • Rochaix, J.D., Malnoë, P.: Anatomy of the chloroplast ribosomal DNA of Chlamydomonas reinhardii. Cell 15, 661–670 (1978)

    Google Scholar 

  • Sanger, F., Air, G.M., Barrell, B.G., Brown, N.L., Coulson, A.R., Fiddes, J.-C., Hutchinson III, C.A., Slocombe, P.M., Smith, M.: Nucleotide sequence of bacteriophage ϕX174 DNA. Nature 265, 687–695 (1977)

    Google Scholar 

  • Schmitz, K.S., Shaw, B.R.: Hydrodynamic evidence in support of spacer regions in chromatin. Science 197, 661–662 (1977)

    Google Scholar 

  • Sharp, P.A., Sugden, B., Sambrook, J.: Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry 12, 3055–3063 (1973)

    Google Scholar 

  • Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–517 (1975)

    Google Scholar 

  • Speirs, J., Birnstiel, M.: Arrangement of the 5.8 S RNA cistrons in the genome of Xenopus laevis. J. Mol. Biol. 87, 237–256 (1974)

    Google Scholar 

  • Sueoka, N., Chiang, K.S., Kates, J.R.: Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardii. 1. Isotopic transfer experiments with a strain producing eight zoospores. J. Mol. Biol. 25, 47–66 (1967)

    Google Scholar 

  • Tewari, K.K., Wildman, S.G.: Function of chloroplast DNA. 1. Hybridization studies involving nuclear and chloroplast DNA with RNA from cytoplasmic (80 S) and chloroplast (70 S) ribosomes. Proc. Natl. Acad. Sci. U.S.A. 59, 569–576 (1968)

    Google Scholar 

  • Thomas, M., Davis, R.W.: Studies on the cleavage of bacteriophage lambda DNA with EcoRI restriction endonuclease. J. Mol. Biol. 91, 315–328 (1975)

    Google Scholar 

  • Vogt, V.M., Braun, R.: Structure of ribosomal DNA in Physarum polycephalum. J. Mol. Biol. 106, 567–587 (1976)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B.: The structural organization of ribosomal DNA in Drosophila melanogaster. Cell 10, 193–212 (1977)

    Google Scholar 

  • 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. 105, 461–486 (1976)

    Google Scholar 

  • White, R.L., Hogness, D.S.: R-loop mapping of the 18 S and 28 S sequences in the long and short repeating units of Drosophila melanogaster DNA. Cell 10, 177–192 (1977)

    Google Scholar 

  • Wilson, G.A., Young, F.E.: Isolation of a sequence specific endonuclease (Bam I) from Bacillus amylolique faciens H. J. Mol. Biol. 97, 123–125 (1975)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Schell

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marco, Y., Rochaix, J.D. Organization of the nuclear ribosomal DNA of Chlamydomonas reinhardii . Molec. Gen. Genet. 177, 715–723 (1980). https://doi.org/10.1007/BF00272684

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00272684

Keywords

Navigation