21st century genetics: Mass spectrometry of yeast telomerase

Kah Wai Lin, Virginia A. Zakian

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

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 RNAto template the addition of telomeric repeats. In addition, one or more accessory proteins are required for telomerase action in vivo. The beststudied 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.

Original languageEnglish (US)
Title of host publication21st Century Genetics Genes at Work, 2015
EditorsTerri Grodzicker, Bruce Stillman, David Stewart
PublisherCold Spring Harbor Laboratory Press
Pages111-116
Number of pages6
ISBN (Print)9781621821472
DOIs
StatePublished - 2016
Event21st Century Genetics Genes at Work, 2015 - Huntington, United States
Duration: May 26 2015May 31 2015

Publication series

NameCold Spring Harbor Symposia on Quantitative Biology
Volume80
ISSN (Print)0091-7451
ISSN (Electronic)1943-4456

Other

Other21st Century Genetics Genes at Work, 2015
CountryUnited States
CityHuntington
Period5/26/155/31/15

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Genetics

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  • Cite this

    Lin, K. W., & Zakian, V. A. (2016). 21st century genetics: Mass spectrometry of yeast telomerase. In T. Grodzicker, B. Stillman, & D. Stewart (Eds.), 21st Century Genetics Genes at Work, 2015 (pp. 111-116). (Cold Spring Harbor Symposia on Quantitative Biology; Vol. 80). Cold Spring Harbor Laboratory Press. https://doi.org/10.1101/sqb.2015.80.027656