Biomedical subjects
Publications and source records attributed to M Teresa Teixeira.
Reconstitution of telomeric DNA at each cell division implies the coordination of DNA semiconservative replication with several processing events still poorly understood. Two reports published recently in Molecular Cell show that a cell-cycle cyclin-dependent kinase, Cdk1p, is required to create the cell-cycle-regulated overhang ().
Telomerase counteracts telomere erosion that stems from incomplete chromosome end replication and nucleolytic processing. A precise understanding of telomere length homeostasis has been hampered by the lack of assays that delineate the nonuniform telomere extension events of single chromosome molecules. Here, we measure telomere elongation at nucleotide resolution in Saccharomyces cerevisiae. The number of nucleotides added to a telomere in a single cell cycle varies between a few to more than 100 nucleotides and is independent of telomere length. Telomerase does not act on every telomere in each cell cycle, however. Instead, it exhibits an increasing preference for telomeres as their lengths decline. Deletion of the telomeric proteins Rif1 or Rif2 gives rise to longer telomeres by increasing the frequency of elongation events. Thus, by taking a molecular snapshot of a single round of telomere replication, we demonstrate that telomere length homeostasis is achieved via a switch between telomerase-extendible and -nonextendible states.
Telomerase extends chromosome ends by iterative reverse transcription of its RNA template. Following the addition of each telomeric repeat, the RNA template and the telomeric substrate reset their relative position in the active site provided by the telomerase reverse transcriptase (TERT). This step might require the formation of guanine-rich secondary structures in the nascent product. Results from numerous studies begin to delineate TERT sub-domains that orchestrate these events and support the model of cooperative action between distinct active sites within telomerase multimers. Natural telomere substrates are protein-DNA complexes that show an asymmetry between the two ends of a chromosome, possibly reflecting their differential mode of replication.
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.