21st Century Genetics: Mass Spectrometry of Yeast Telomerase. Author Kah Lin, Virginia Zakian Publication Year 2015 Type Journal Article Abstract Telomerase is a specialized reverse transcriptase that maintains the ends of chromosomes in almost all eukaryotes. The core of telomerase consists of telomerase RNA and the reverse transcriptase that uses a short segment without the RNA to template the addition of telomeric repeats. In addition, one or more accessory proteins are required for telomerase action in vivo. The best-studied accessory protein is Est1, which is conserved from yeasts to humans. In budding yeast, Est1 has two critical in vivo functions: By interaction with Cdc13, a telomere-binding protein, it recruits telomerase to telomeres, and it also increases telomerase activity. Although budding yeast telomerase is highly regulated by the cell cycle, Est1 is the only telomerase subunit whose abundance is cell cycle-regulated. Close to 400 yeast genes are reported to affect telomere length, although the specific function of most of them is unknown. With the goal of identifying novel telomerase regulators by mass spectrometry, we developed methods for purifying yeast telomerase and its associated proteins. We summarize the methods we used and describe the experiments that show that four telomerase-associated proteins identified by mass spectrometry, none of which had been linked previously to telomeres, affect telomere length and cell cycle regulation of telomerase by controlling Est1 abundance. Keywords Mass Spectrometry, Telomerase, Telomere-Binding Proteins, Fungal Proteins, Yeasts, G1 Phase Cell Cycle Checkpoints, G2 Phase Cell Cycle Checkpoints, Valosin Containing Protein Journal Cold Spring Harb Symp Quant Biol Volume 80 Pages 111-6 Date Published 2015 ISSN Number 1943-4456 DOI 10.1101/sqb.2015.80.027656 Alternate Journal Cold Spring Harb Symp Quant Biol PMCID PMC5441543 PMID 26763982 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML