Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1985 Aug 26;13(16):5817–5831. doi: 10.1093/nar/13.16.5817

Macronuclear DNA of Tetrahymena thermophila exists as defined subchromosomal-sized molecules.

M I Altschuler, M C Yao
PMCID: PMC321914  PMID: 4034396

Abstract

Using the method of orthogonal-field-alternation gel electrophoresis, we have resolved the macronuclear DNA of Tetrahymena thermophila into a series of distinct bands. Using electrode switching intervals ranging from 10 to 70 seconds we have resolved DNA bands ranging in size from about 21 kb up to and beyond the size of yeast chromosomes VII and XV. Hybridization of Southern blots from these gels to both unique and repetitive DNA sequences shows that the macronuclear genome of T. thermophila has a precise organization. The unique sequences tested each hybridize to only one band of macronuclear DNA and the hybridization patterns seem to be identical in several inbred strains examined.

Full text

PDF
5817

Images in this article

5820Image
on p.5820
5821Image
on p.5821
5822Image
on p.5822
5824Image
on p.5824
5825Image
on p.5825
5826Image
on p.5826
5827Image
on p.5827

Selected References

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

  1. Allen S. L., Ervin P. R., McLaren N. C., Brand R. E. The 5S ribosomal RNA gene clusters in Tetrahymena thermophila: strain differences, chromosomal localization, and loss during micronuclear ageing. Mol Gen Genet. 1984;197(2):244–253. doi: 10.1007/BF00330970. [DOI] [PubMed] [Google Scholar]
  2. Austerberry C. F., Allis C. D., Yao M. C. Specific DNA rearrangements in synchronously developing nuclei of Tetrahymena. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7383–7387. doi: 10.1073/pnas.81.23.7383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bannon G. A., Bowen J. K., Yao M. C., Gorovsky M. A. Tetrahymena H4 genes: structure, evolution and organization in macro- and micronuclei. Nucleic Acids Res. 1984 Feb 24;12(4):1961–1975. doi: 10.1093/nar/12.4.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bruns P. J., Brussard T. E. Nullisomic tetrahymena: eliminating germinal chromosomes. Science. 1981 Jul 31;213(4507):549–551. doi: 10.1126/science.213.4507.549. [DOI] [PubMed] [Google Scholar]
  5. Callahan R. C., Shalke G., Gorovsky M. A. Developmental rearrangements associated with a single type of expressed alpha-tubulin gene in Tetrahymena. Cell. 1984 Feb;36(2):441–445. doi: 10.1016/0092-8674(84)90237-x. [DOI] [PubMed] [Google Scholar]
  6. Carle G. F., Olson M. V. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. doi: 10.1093/nar/12.14.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  8. Gorovsky M. A. Genome organization and reorganization in Tetrahymena. Annu Rev Genet. 1980;14:203–239. doi: 10.1146/annurev.ge.14.120180.001223. [DOI] [PubMed] [Google Scholar]
  9. Jackson D. A., Cook P. R. A general method for preparing chromatin containing intact DNA. EMBO J. 1985 Apr;4(4):913–918. doi: 10.1002/j.1460-2075.1985.tb03718.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Katzen A. L., Cann G. M., Blackburn E. H. Sequence-specific fragmentation of macronuclear DNA in a holotrichous ciliate. Cell. 1981 May;24(2):313–320. doi: 10.1016/0092-8674(81)90321-4. [DOI] [PubMed] [Google Scholar]
  12. Kimmel A. R., Gorovsky M. A. Organization of the 5S RNA genes in macro- and micronuclei of Tetrahymena pyriformis. Chromosoma. 1978 Jun 23;67(1):1–20. doi: 10.1007/BF00285644. [DOI] [PubMed] [Google Scholar]
  13. King B. O., Yao M. C. Tandemly repeated hexanucleotide at Tetrahymena rDNA free end is generated from a single copy during development. Cell. 1982 Nov;31(1):177–182. doi: 10.1016/0092-8674(82)90417-2. [DOI] [PubMed] [Google Scholar]
  14. Klobutcher L. A., Swanton M. T., Donini P., Prescott D. M. All gene-sized DNA molecules in four species of hypotrichs have the same terminal sequence and an unusual 3' terminus. Proc Natl Acad Sci U S A. 1981 May;78(5):3015–3019. doi: 10.1073/pnas.78.5.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mortimer R. K., Schild D. Genetic map of Saccharomyces cerevisiae. Microbiol Rev. 1980 Dec;44(4):519–571. doi: 10.1128/mr.44.4.519-571.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pan W. C., Blackburn E. H. Single extrachromosomal ribosomal RNA gene copies are synthesized during amplification of the rDNA in Tetrahymena. Cell. 1981 Feb;23(2):459–466. doi: 10.1016/0092-8674(81)90141-0. [DOI] [PubMed] [Google Scholar]
  17. Pederson D. S., Yao M. C., Kimmel A. R., Gorovsky M. A. Sequence organization within and flanking clusters of 5S ribosomal RNA genes in Tetrahymena. Nucleic Acids Res. 1984 Mar 26;12(6):3003–3021. doi: 10.1093/nar/12.6.3003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  19. Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
  20. Schwartz D. C., Saffran W., Welsh J., Haas R., Goldenberg M., Cantor C. R. New techniques for purifying large DNAs and studying their properties and packaging. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):189–195. doi: 10.1101/sqb.1983.047.01.024. [DOI] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Swanton M. T., Greslin A. F., Prescott D. M. Arrangement of coding and non-coding sequences in the DNA molecules coding for rRNAs in Oxytricha sp. DNA of ciliated protozoa. VII. Chromosoma. 1980;77(2):203–215. doi: 10.1007/BF00329545. [DOI] [PubMed] [Google Scholar]
  23. Swanton M. T., Heumann J. M., Prescott D. M. Gene-sized DNA molecules of the macronuclei in three species of hypotrichs: size distributions and absence of nicks. DNA of ciliated protozoa. VIII. Chromosoma. 1980;77(2):217–227. doi: 10.1007/BF00329546. [DOI] [PubMed] [Google Scholar]
  24. Weindruch R. H., Doerder F. P. Age-dependent micronuclear deterioration in Tetrahymena pyriformis, syngen 1. Mech Ageing Dev. 1975 May-Aug;4(3-4):263–279. doi: 10.1016/0047-6374(75)90028-7. [DOI] [PubMed] [Google Scholar]
  25. Williams J. B., Fleck E. W., Hellier L. E., Uhlenhopp E. Viscoelastic studies on Tetrahymena macronuclear DNA. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5062–5065. doi: 10.1073/pnas.75.10.5062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Woodard J., Kaneshiro E., Gorovsky M. A. Cytochemical studies on the problem of macronuclear subnuclei in tetrahymena. Genetics. 1972 Feb;70(2):251–260. doi: 10.1093/genetics/70.2.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yao M. C., Blackburn E., Gall J. Tandemly repeated C-C-C-C-A-A hexanucleotide of Tetrahymena rDNA is present elsewhere in the genome and may be related to the alteration of the somatic genome. J Cell Biol. 1981 Aug;90(2):515–520. doi: 10.1083/jcb.90.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yao M. C., Choi J., Yokoyama S., Austerberry C. F., Yao C. H. DNA elimination in Tetrahymena: a developmental process involving extensive breakage and rejoining of DNA at defined sites. Cell. 1984 Feb;36(2):433–440. doi: 10.1016/0092-8674(84)90236-8. [DOI] [PubMed] [Google Scholar]
  29. Yao M. C. Elimination of specific DNA sequences from the somatic nucleus of the ciliate Tetrahymena. J Cell Biol. 1982 Mar;92(3):783–789. doi: 10.1083/jcb.92.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yao M. C., Gorovsky M. A. Comparison of the sequences of macro- and micronuclear DNA of Tetrahymena pyriformis. Chromosoma. 1974;48(1):1–18. doi: 10.1007/BF00284863. [DOI] [PubMed] [Google Scholar]
  31. Yao M. C. Ribosomal RNA gene amplification in Tetrahymena may be associated with chromosome breakage and DNA elimination. Cell. 1981 Jun;24(3):765–774. doi: 10.1016/0092-8674(81)90102-1. [DOI] [PubMed] [Google Scholar]
  32. Yao M. C., Yao C. H. Repeated hexanucleotide C-C-C-C-A-A is present near free ends of macronuclear DNA of Tetrahymena. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7436–7439. doi: 10.1073/pnas.78.12.7436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yokoyama R., Yao M. C. Internal micronuclear DNA regions which include sequences homologous to macronuclear telomeres are deleted during development in Tetrahymena. Nucleic Acids Res. 1984 Aug 10;12(15):6103–6116. doi: 10.1093/nar/12.15.6103. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES