RNA, Ribosomal

Publication Title: 
The Journal of Protozoology

We have analyzed the macronuclear DNA of Paramecium tetraurelia using orthogonal-field-alternation gel electrophoresis. The mean size of the linear macronuclear DNA molecules is approximately 450 kb. Less than 6% of the macronuclear DNA is larger than 800 kb. Using pulse times of 20, 40, 60 and 90 s we show that the macronuclear fragment containing the A type variable surface antigen gene migrates reproducibly as a 320-kb linear DNA. Over the same pulse times we describe the unusual migration of the ribosomal RNA gene (rDNA) of P. tetraurelia.

Author(s): 
Phan, H. L.
Forney, J.
Blackburn, E. H.
Publication Title: 
Cell

We have analyzed the de novo telomere synthesis catalyzed by the enzyme telomere terminal transferase (telomerase) from Tetrahymena. Oligonucleotides representing the G-rich strand of telomeric sequences from five different organisms specifically primed the addition of TTGGGG repeats in vitro, suggesting that primer recognition may involve a DNA structure unique to these oligonucleotides. The sequence at the 3' end of the oligonucleotide primer specified the first nucleotide added in the reaction.

Author(s): 
Greider, C. W.
Blackburn, E. H.
Publication Title: 
The Journal of Biological Chemistry

We have determined the complete sequence of the nuclear gene encoding the small subunit (17 S) rRNA of the ciliated protozoan Tetrahymena thermophila. The gene encodes an RNA molecule which is 1753 nucleotides in length. The sequence of the Tetrahymena small subunit rRNA is homologous to those of other eukaryotes, and the predicted secondary structure for the molecule includes features which are characteristic of eukaryotic small subunit rRNAs.

Author(s): 
Spangler, E. A.
Blackburn, E. H.
Publication Title: 
Nucleic Acids Research

The rRNA genes in the somatic macronucleus of Tetrahymena thermophila are normally on 21 kb linear palindromic molecules (rDNA). We examined the effect on rRNA gene dosage of transforming T.thermophila macronuclei with plasmid constructs containing a pair of tandemly repeated rDNA replication origin regions unlinked to the rRNA gene. A significant proportion of the plasmid sequences were maintained as high copy circular molecules, eventually consisting solely of tandem arrays of origin regions.

Author(s): 
Pan, W. J.
Blackburn, E. H.
Publication Title: 
Cell

The amplification of ribosomal DNA during development of the somatic macronucleus in Tetrahymena thermophila was analyzed by genetic and molecular biological techniques. We have identified an alternate form of the rDNA, structurally distinguishable from the wild-type by an extra cutting site for Bam HI in its nontranscribed spacer. The altered rDNA was inherited in crosses in a simple Mendelian fashion, consistent with the presence of only one rRNA gene copy per haploid genome in the micronucleus.

Author(s): 
Pan, W. C.
Orias, E.
Flacks, M.
Blackburn, E. H.
Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

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.

Author(s): 
Blackburn, E. H.
Chiou, S. S.
Publication Title: 
The Journal of Cell Biology

The ribosomal RNA genes of the Tetrahymena macronucleus exist as extrachromosomal, linear molecules. The termini of these molecules have been shown to contain the tandemly repeated hexanucleotide (C-C-C-C-A-A)n. In this study the same or related sequences were found in other locations of the genome. Using the depurination method, we showed that macronuclear DNA contained this sequence even after rDNA had been removed. The sequence was found mainly in the repetitive fraction of the DNA.

Author(s): 
Yao, M. C.
Blackburn, E.
Gall, J.
Publication Title: 
Cell

Macronuclear DNA from the protozoan G. chattoni, a holotrichous ciliate, was analyzed. Most, if not all, of the macronuclear DNA is subchromosomal, ranging in size from above 100 kb down to 2.1 kb, with molecules in the lower molecular weight range being resolvable by gel electrophoresis into reproducible, specific, discrete size classes. A prominent class of linear 9.3 kb molecules consists of single free rRNA genes. Upon denaturation of total macronuclear DNA was found as single-stranded circles.

Author(s): 
Katzen, A. L.
Cann, G. M.
Blackburn, E. H.
Publication Title: 
The Journal of Cell Biology

The ribosomal RNA genes of the Tetrahymena macronucleus exist as extrachromosomal, linear molecules. The termini of these molecules have been shown to contain the tandemly repeated hexanucleotide (C-C-C-C-A-A)n. In this study the same or related sequences were found in other locations of the genome. Using the depurination method, we showed that macronuclear DNA contained this sequence even after rDNA had been removed. The sequence was found mainly in the repetitive fraction of the DNA.

Author(s): 
Yao, M. C.
Blackburn, E.
Gall, J.
Publication Title: 
Cell

A novel form of extrachromosomal rDNA has been identified in conjugating Tetrahymena cells. This rDNA consists of 11 kb linear double-stranded DNA molecules, each containing a single rRNA gene copy. The DNA sequence, tandemly repeated CCCCAA (Blackburn and Gall, 1978) found at the termini of extrachromosomal palindromic rDNA (the macronuclear form found in vegetatively growing cells), is also present at the corresponding terminus of the 11 kb rDNA.

Author(s): 
Pan, W. C.
Blackburn, E. H.

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