Non-nucleosomal Packaging of a Tandemly Repeated DNA Sequence at Termini of Extrachromosomal DNA Coding for RRNA in Tetrahymena

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1981 Apr 1

PMID 6941283

Citations 28

Authors

E H Blackburn

S S Chiou

Affiliations

Soon will be listed here.

Abstract

A tandemly repeated DNA hexanucleotide sequence, 5'C-C-C-C-A-A3', that occurs at the termini of extrachromosomal DNA molecules coding for rRNA (rDNA) in Tetrahymena macronuclei was examined to determine whether it is packaged as nucleosomes. This repeated DNA sequence comprises the terminal few hundred base pairs at each end of the linear rDNA molecules. Digestion of macronuclei with micrococcal nuclease showed that this DNA sequence is protected from digestion but is left, following digestion, as a single but broad size class of DNA fragments several hundred base pairs long, under conditions in which bulk macronuclear DNA and rDNA were digested to fragments that were multiples of approximately 200 base pairs in length. The repeated C-C-C-C-A-A was found protected as fragments longer than the bulk macronuclear DNA digestion products at all times during digestion. Together with putative associated protein(s), this protected DNA was soluble after lysis of micrococcal nuclease-digested macronuclei at low salt concentrations but was insoluble in 0.075--0.2 M KCl, regardless of the extend of digestion. The size and solubility properties of the repeated C-C-C-C-A-A DNA nucleoprotein complex after micrococcal nuclease digestion of macronuclei are clearly distinguishable from those of nucleosomes, and it is inferred that this DNA sequence in macronuclei is packaged in chromatin by proteins other than histones.

Citing Articles

Aberrant gene expression profiles, during in vitro osteoblast differentiation, of telomerase deficient mouse bone marrow stromal stem cells (mBMSCs).

Saeed H, Iqtedar M J Biomed Sci. 2015; 22:11.

PMID: 25633569 PMC: 4318164. DOI: 10.1186/s12929-015-0116-4.

Stem cell function and maintenance - ends that matter: role of telomeres and telomerase.

Saeed H, Iqtedar M J Biosci. 2013; 38(3):641-9.

PMID: 23938394 DOI: 10.1007/s12038-013-9346-3.

The Pot1a-associated proteins Tpt1 and Pat1 coordinate telomere protection and length regulation in Tetrahymena.

Linger B, Morin G, Price C Mol Biol Cell. 2011; 22(21):4161-70.

PMID: 21900503 PMC: 3204076. DOI: 10.1091/mbc.E11-06-0551.

Characterization of the yeast telomere nucleoprotein core: Rap1 binds independently to each recognition site.

Williams T, Levy D, Maki-Yonekura S, Yonekura K, Blackburn E J Biol Chem. 2010; 285(46):35814-24.

PMID: 20826803 PMC: 2975205. DOI: 10.1074/jbc.M110.170167.

Modulation of telomere length dynamics by the subtelomeric region of tetrahymena telomeres.

Jacob N, Stout A, Price C Mol Biol Cell. 2004; 15(8):3719-28.

PMID: 15169872 PMC: 491831. DOI: 10.1091/mbc.e04-03-0237.

References

Kornberg R . Structure of chromatin. Annu Rev Biochem. 1977; 46:931-54. DOI: 10.1146/annurev.bi.46.070177.004435. View

Blackburn E, Gall J . A tandemly repeated sequence at the termini of the extrachromosomal ribosomal RNA genes in Tetrahymena. J Mol Biol. 1978; 120(1):33-53. DOI: 10.1016/0022-2836(78)90294-2. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Wild M, Gall J . An intervening sequence in the gene coding for 25S ribosomal RNA of Tetrahymena pigmentosa. Cell. 1979; 16(3):565-73. DOI: 10.1016/0092-8674(79)90030-8. View

Noll M, Kornberg R . Action of micrococcal nuclease on chromatin and the location of histone H1. J Mol Biol. 1977; 109(3):393-404. DOI: 10.1016/s0022-2836(77)80019-3. View

Sedat J, Manuelidis L . A direct approach to the structure of eukaryotic chromosomes. Cold Spring Harb Symp Quant Biol. 1978; 42 Pt 1:331-50. DOI: 10.1101/sqb.1978.042.01.035. View

Yao M, Blackburn E, Gall J . Amplification of the rRNA genes in Tetrahymena. Cold Spring Harb Symp Quant Biol. 1979; 43 Pt 2:1293-6. DOI: 10.1101/sqb.1979.043.01.147. View

Noll M, Thomas J, Kornberg R . Preparation of native chromatin and damage caused by shearing. Science. 1975; 187(4182):1203-6. DOI: 10.1126/science.187.4182.1203. View

Groudine M, Das S, Neiman P, Weintraub H . Regulation of expression and chromosomal subunit conformation of avian retrovirus genomes. Cell. 1978; 14(4):865-78. DOI: 10.1016/0092-8674(78)90342-2. View

10.

Maniatis T, Jeffrey A, Kleid D . Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975; 72(3):1184-8. PMC: 432491. DOI: 10.1073/pnas.72.3.1184. View

11.

Jones R . Preparation of chromatin containing ribosomal deoxyribonucleic acid from the macronucleus of Tetrahymena pyriformis. Biochem J. 1978; 173(1):145-53. PMC: 1185757. DOI: 10.1042/bj1730145. View

12.

Engelke D, Ng S, Shastry B, Roeder R . Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes. Cell. 1980; 19(3):717-28. DOI: 10.1016/s0092-8674(80)80048-1. View

13.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

14.

McCarty Jr K, Vollmer R, McCarty K . Improved computer program data for the resolution and fractionation of macromolecules by isokinetic sucrose density gradient sedimentation. Anal Biochem. 1974; 61(1):165-83. DOI: 10.1016/0003-2697(74)90343-1. View

15.

Botchan M, Topp W, Sambrook J . The arrangement of simian virus 40 sequences in the DNA of transformed cells. Cell. 1976; 9(2):269-87. DOI: 10.1016/0092-8674(76)90118-5. View

16.

Levinger L, Carter Jr C . Superstructural differences between chromatin in nuclei and in solution are revealed by kinetics of micrococcal nuclease digestion. J Biol Chem. 1979; 254(19):9477-87. View

17.

Gorovsky M, Glover C, Johmann C, Keevert J, Mathis D, Samuelson M . Histones and chromatin structure in Tetrahymena macro- and micronuclei. Cold Spring Harb Symp Quant Biol. 1978; 42 Pt 1:493-503. DOI: 10.1101/sqb.1978.042.01.052. View

18.

Olins A, Carlson R, WRIGHT E, Olins D . Chromatin nu bodies: isolation, subfractionation and physical characterization. Nucleic Acids Res. 1976; 3(12):3271-91. PMC: 343173. DOI: 10.1093/nar/3.12.3271. View

19.

Allis C, Glover C, Gorovsky M . Micronuclei of Tetrahymena contain two types of histone H3. Proc Natl Acad Sci U S A. 1979; 76(10):4857-61. PMC: 413036. DOI: 10.1073/pnas.76.10.4857. View

20.

BLUMENFELD M, Orf J, Sina B, Kreber R, Callahan M, Mullins J . Correlation between phosphorylated H1 histones and satellite DNAs in Drosophila virilis. Proc Natl Acad Sci U S A. 1978; 75(2):866-70. PMC: 411358. DOI: 10.1073/pnas.75.2.866. View