RNA, Protozoan

Publication Title: 
Cold Spring Harbor Symposia on Quantitative Biology
Author(s): 
Lee, M. S.
Gallagher, R. C.
Bradley, J.
Blackburn, E. H.
Publication Title: 
The EMBO journal

Telomerase, an essential ribonucleoprotein reverse transcriptase, adds telomeric DNA to the ends of eukaryotic chromosomes. We examined the conformational properties of the naked RNA moiety of telomerase from two related ciliates, Tetrahymena thermophila and Glaucoma chattoni. As well as finding evidence for features proposed previously on the basis of phylogenetic comparisons, novel conserved structural properties were revealed.

Author(s): 
Bhattacharyya, A.
Blackburn, E. H.
Publication Title: 
Annual Review of Biochemistry
Author(s): 
Blackburn, E. H.
Publication Title: 
Molecular and Cellular Biology

The ribonucleoprotein enzyme telomerase is a specialized reverse transcriptase that synthesizes telomeric DNA by copying a template sequence within the telomerase RNA. Here we analyze the actions of telomerase from Tetrahymena thermophila assembled in vivo with mutated or wild-type telomerase RNA to define further the roles of particular telomerase RNA residues involved in essential enzymatic functions: templating, substrate alignment, and promotion of polymerization.

Author(s): 
Gilley, D.
Blackburn, E. H.
Publication Title: 
Genes & Development

The ribonucleoprotein enzyme telomerase synthesizes telomeric DNA by copying a template sequence in the telomerase RNA. We studied the functional roles of specific residues in the Tetrahymena telomerase RNA template region. Unexpectedly, mutation of certain templating residues caused dramatic effects on specific aspects of the enzyme reaction, including loss of enzymatic fidelity and premature product dissociation. None of these fundamental changes in enzymatic action are explainable by altered base-pairing between the telomerase RNA and DNA substrate.

Author(s): 
Gilley, D.
Lee, M. S.
Blackburn, E. H.
Publication Title: 
Science (New York, N.Y.)

Telomeres are essential for chromosome stability, but their functions at specific cell-cycle stages are unknown. Telomeres are now shown to have a role in chromosome separation during mitosis. In telomeric DNA mutants of Tetrahymena thermophila, created by expression of a telomerase RNA with an altered template sequence, division of the germline nucleus was severely delayed or blocked in anaphase. The mutant chromatids failed to separate completely at the midzone, becoming stretched to up to twice their normal length. These results suggest a physical block in mutant telomere separation.

Author(s): 
Kirk, K. E.
Harmon, B. P.
Reichardt, I. K.
Sedat, J. W.
Blackburn, E. H.
Publication Title: 
The EMBO journal

Telomerase is a reverse transcriptase minimally composed of a reverse transcriptase protein subunit and an internal RNA component that contains the templating region. Point mutations of template RNA bases can cause loss of enzymatic activity, reduced processivity and misincorporation in vitro. Here we report the first complete replacement of the nine base TETRAHYMENA: thermophila telomerase templating region in vivo with non-telomeric sequences. Rather than ablating telomerase activity, three such replaced telomerases (U9, AUN and AU4) were effective in polymerization in vitro.

Author(s): 
Ware, T. L.
Wang, H.
Blackburn, E. H.
Publication Title: 
Genes & Development

During development of the somatic macronucleus of Tetrahymena thermophila, the rDNA is excised from its germ-line chromosome, rearranged into a palindrome, and amplified to 10(4) copies. We have identified a cis-acting germ-line mutation, rmm11/6, that prevents amplification of the rDNA in all but approximately 1 in 10(5) cells when it is the only rDNA allele in the developing macronucleus. The rmm11/6 mutation resides in a conserved element required for excision, the chromosome breakage sequence (Cbs) flanking the 3' end of the rDNA.

Author(s): 
Kapler, G. M.
Blackburn, E. H.
Publication Title: 
Methods in Cell Biology
Author(s): 
Blackburn, E.
Gilley, D.
Ware, T.
Bhattacharyya, A.
Kirk, K.
Wang, H.
Publication Title: 
Proceedings of the National Academy of Sciences of the United States of America

The ribonucleoprotein (RNP) enzyme telomerase is required for replication of eukaryotic chromosomal termini. The RNA moiety of telomerase is essential for enzyme function and provides the template for telomeric DNA synthesis. However, the roles of its nontemplate domains have not been explored. Here we demonstrate that a novel interspecies telomerase RNA swap in vivo creates a functional but aberrant telomerase. Telomerase RNA from the ciliate Glaucoma chattoni was expressed in Tetrahymena thermophila cells.

Author(s): 
Bhattacharyya, A.
Blackburn, E. H.

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