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CST does not evict elongating telomerase but prevents initiation by ssDNA binding

Zaug, Arthur J. and Lim, Ci Ji and Olson, Conner L. and Carilli, Maria T. and Goodrich, Karen J. and Wuttke, Deborah S. and Cech, Thomas R. (2021) CST does not evict elongating telomerase but prevents initiation by ssDNA binding. Nucleic Acids Research, 49 (20). pp. 11653-11665. ISSN 0305-1048. PMCID PMC8599947. doi:10.1093/nar/gkab942. https://resolver.caltech.edu/CaltechAUTHORS:20210831-220545152

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Abstract

The CST complex (CTC1-STN1-TEN1) has been shown to inhibit telomerase extension of the G-strand of telomeres and facilitate the switch to C-strand synthesis by DNA polymerase alpha-primase (pol α-primase). Recently the structure of human CST was solved by cryo-EM, allowing the design of mutant proteins defective in telomeric ssDNA binding and prompting the reexamination of CST inhibition of telomerase. The previous proposal that human CST inhibits telomerase by sequestration of the DNA primer was tested with a series of DNA-binding mutants of CST and modeled by a competitive binding simulation. The DNA-binding mutants had substantially reduced ability to inhibit telomerase, as predicted from their reduced affinity for telomeric DNA. These results provide strong support for the previous primer sequestration model. We then tested whether addition of CST to an ongoing processive telomerase reaction would terminate DNA extension. Pulse-chase telomerase reactions with addition of either wild-type CST or DNA-binding mutants showed that CST has no detectable ability to terminate ongoing telomerase extension in vitro. The same lack of inhibition was observed with or without pol α-primase bound to CST. These results suggest how the switch from telomerase extension to C-strand synthesis may occur.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1093/nar/gkab942DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc8599947/PubMed CentralArticle
https://github.com/mtcarilli/CST_Inhibits_TelomeraseRelated ItemCode
https://doi.org/10.1101/2021.08.25.457677DOIDiscussion Paper
ORCID:
AuthorORCID
Zaug, Arthur J.0000-0001-7324-803X
Lim, Ci Ji0000-0003-3327-1926
Goodrich, Karen J.0000-0003-3301-7725
Wuttke, Deborah S.0000-0002-8158-8795
Cech, Thomas R.0000-0001-7338-3389
Additional Information:© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. Received August 25, 2021; Revised September 29, 2021; Editorial Decision September 29, 2021; Accepted October 01, 2021. We thank D. Herschlag (Stanford U.) and J. Lingner (EPFL, Lausanne) for valuable discussions. We thank T. Nahreini for her excellent management of the Biochemistry Cell Culture Facility, CU Boulder. T.R.C. is an investigator of the Howard Hughes Medical Institute. Funding: National Institutes of Health [R00 GM131023 to C.L., R01 GM139274 to D.S.W.]; National Science Foundation [MCB 1716425 to D.S.W.]; M.T.C. was supported by the Biological Sciences Initiative funded by the University of Colorado Boulder; Howard Hughes Medical Institute through the Science Education Program. Funding for open access charge: Howard Hughes Medical Institute and the University of Colorado Boulder Open Access Fund. Conflict of interest statement. T.R.C. is a scientific advisor for Storm Therapeutics and Eikon Therapeutics.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIHR00 GM131023
NIHR01 GM139274
NSFMCB-1716425
University of Colorado, BoulderUNSPECIFIED
Issue or Number:20
PubMed Central ID:PMC8599947
DOI:10.1093/nar/gkab942
Record Number:CaltechAUTHORS:20210831-220545152
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210831-220545152
Official Citation:Arthur J Zaug, Ci Ji Lim, Conner L Olson, Maria T Carilli, Karen J Goodrich, Deborah S Wuttke, Thomas R Cech, CST does not evict elongating telomerase but prevents initiation by ssDNA binding, Nucleic Acids Research, Volume 49, Issue 20, 18 November 2021, Pages 11653–11665, https://doi.org/10.1093/nar/gkab942
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:110674
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:31 Aug 2021 22:14
Last Modified:03 Jan 2022 22:35

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