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Intracellular trafficking of yeast telomerase components

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TLDR
It is found that Est1p, Est2p and TLC1 can migrate independently of each other to the nucleus and a role of the nucleolus in telomerase biogenesis is suggested.
Abstract
Telomerase uses an internal RNA moiety as template for the synthesis of telomere repeats. In Saccharomyces cerevisiae, the telomerase holoenzyme contains the telomerase reverse transcriptase subunit Est2p, the telomerase RNA moiety TLC1, the telomerase associated proteins Est1p and Est3p, and Sm proteins. Here we assess telomerase assembly by determining the localization of telomerase components. We found that Est1p, Est2p and TLC1 can migrate independently of each other to the nucleus. With limiting amounts of TLC1, overexpressed Est1p and Est2p accumulated in the nucleolus, whereas enzymatically active Est2p–TLC1 complexes are distributed over the entire nucleus. The distribution to the nucleoplasm depended on the specific interaction between Est2p and TLC1 but was independent of Est1p and Est3p. Altogether, our results suggest a role of the nucleolus in telomerase biogenesis. We also describe experiments that support a transient cytoplasmic localization of TLC1 RNA.

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Journal ArticleDOI

Cellular dynamics of small RNAs.

TL;DR: Data that support or argue against nucleus/cytoplasm bidirectional movement for each category of small RNA and the possible roles that such movement may serve are discussed.
Journal ArticleDOI

AtTRB1, a telomeric DNA-binding protein from Arabidopsis, is concentrated in the nucleolus and shows highly dynamic association with chromatin.

TL;DR: The results reveal that AtTRB1 interaction with chromatin is regulated at two levels at least, one of which is coupled with cell-cycle progression, with the other involving rapid exchange.
Journal ArticleDOI

Characterization of recombinant Saccharomyces cerevisiae telomerase core enzyme purified from yeast

TL;DR: The procedure of reconstitution and purification of telomerase core enzyme that is developed now allows for further mechanistic studies of the functions of other subunits of the telomersase holoenzyme as well as other telomer enzyme regulation proteins.
Journal ArticleDOI

Regulation of telomere length by an N-terminal region of the yeast telomerase reverse transcriptase

TL;DR: The identification of mutations within an N-terminal region (region I) of the yeast telomerase catalytic subunit (Est2p) that cause telomere lengthening without altering measurable catalytic properties of the enzyme in vitro are reported.
References
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Journal ArticleDOI

A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis.

TL;DR: The essential RNA component of this ribonucleoprotein enzyme has now been cloned and found to contain the sequence CAACCCCAA, which seems to be the template for the synthesis of TTGGGG repeats.
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Reverse Transcriptase Motifs in the Catalytic Subunit of Telomerase

TL;DR: The reverse transcriptase protein fold, previously known to be involved in retroviral replication and retrotransposition, is essential for normal chromosome telomere replication in diverse eukaryotes.
Journal ArticleDOI

A telomerase component is defective in the human disease dyskeratosis congenita

TL;DR: It is found that primary fibroblasts and lymphoblasts from DKC-affected males are not detectably deficient in conventional H/ACA small nucleolar RNA accumulation or function; however, DKC cells have a lower level of telomerase RNA, produce lower levels of telomersase activity and have shorter telomeres than matched normal cells.
Journal ArticleDOI

A mutant with a defect in telomere elongation leads to senescence in yeast.

TL;DR: Using this assay, a mutant that displays a progressive decrease in telomere length as well as an increased frequency of chromosome loss is isolated, which defines a new gene, designated EST1 (for ever shorter telomeres).
Journal ArticleDOI

Ribosome synthesis in Saccharomyces cerevisiae.

TL;DR: The recent, and often surprising, advances in the understanding of ribosome synthesis in the yeast Saccharomyces cerevisiae will underscore the unexpected complexity of eukaryotic ribosomes synthesis.
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