Cryo-EM of human telomerase — ASN Events

Cryo-EM of human telomerase (#211)

Scott Cohen 1 , Rosalba Rothnagel 2 , George Lovrecz 3 , Tracy Bryan 1 , Michael Parker 4 , Ben Hankamer 2
  1. Children's Medical Research Institute, Westmead, NSW, Australia
  2. Institute for Molecular Bioscience, University of Queensland, St. Lucia, QLD, Australia
  3. CSIRO Biomedical Manufacturing, Clayton, VIC, Australia
  4. St. Vincent's Institute for Medical Research, Fitzroy, VIC, Australia

Telomeres, the repetitive DNA-protein complexes at the ends of linear chromosomes, shorten with each cycle of DNA replication, providing a counting mechanism to limit the number of times a cell can divide. Most cancer cells have activated the ribonucleoprotein enzyme telomerase to add telomeric DNA repeats and counteract telomere shortening, allowing for unlimited proliferation. In contrast, normal cells have undetectable or low levels of telomerase. Inhibition of telomerase is a promising approach for cancer therapy that should be effective against a broad range of cancers while displaying few side effects.

 

We reported the purification and composition of the human telomerase enzyme complex, consisting of two molecules each of: i) the telomerase reverse transcriptase catalytic protein component; ii) the telomerase RNA; and iii) the RNA-binding protein dyskerin (1). Building on this knowledge we established an over-expression system in suspension HEK-293T cells that yields ~400-fold greater activity over endogenous levels, performed on the 20-Litre scale; this system is providing sufficient telomerase for cryo-electron microscopy. Progress on data (particle) collection and processing will be presented.

 

(1) Cohen SB, et al. (2007) Science, 315, pp 1850-1853.

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