Telomere dynamics in cells with introduced telomerase: A rapid assay for telomerase activity on telomeres

Patricia A. McChesney, Dara L. Aisner, Bryan C. Frank, Woodring E. Wright, Jerry W. Shay

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Most immortal cell lines derived from human cancers or transformed in vitro maintain telomeres by endogenous expression of telomerase. In the present work, immortal cells that already express endogenous telomerase activity were induced to overexpress an exogenous telomerase (hTERT) and were analyzed for changes in telomere length. Introduction of hTERT into telomerase-positive immortal cell lines results in elevated telomerase activity as measured by the TRAP assay, leading to elongated telomeres in the cell lines tested. We explore possibilities for regulatory differences among the cell lines, including the level of over-expression of the catalytic subunit hTERT and the endogenous levels of telomere binding proteins. Reducing levels of hTERT expression with a construct containing an inefficient translation initiation sequence provided sufficient telomerase expression for maximal rates of telomere elongation. Overexpression of the hTERT alters the telomere length normally maintained in these cells and provides a very useful assay for the rapid analysis of telomerase activity on its native substrate, telomeres. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)312-318
Number of pages7
JournalMolecular Cell Biology Research Communications
Volume3
Issue number5
DOIs
StatePublished - 2000

Fingerprint

Telomerase
Telomere
Cell Line
Telomere-Binding Proteins
Catalytic Domain
Neoplasms

Keywords

  • hTERT
  • hTR
  • Telomerase
  • Telomere
  • TRAP

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Telomere dynamics in cells with introduced telomerase : A rapid assay for telomerase activity on telomeres. / McChesney, Patricia A.; Aisner, Dara L.; Frank, Bryan C.; Wright, Woodring E.; Shay, Jerry W.

In: Molecular Cell Biology Research Communications, Vol. 3, No. 5, 2000, p. 312-318.

Research output: Contribution to journalArticle

McChesney, Patricia A. ; Aisner, Dara L. ; Frank, Bryan C. ; Wright, Woodring E. ; Shay, Jerry W. / Telomere dynamics in cells with introduced telomerase : A rapid assay for telomerase activity on telomeres. In: Molecular Cell Biology Research Communications. 2000 ; Vol. 3, No. 5. pp. 312-318.
@article{0bbad8d051344756a21ad011063588fa,
title = "Telomere dynamics in cells with introduced telomerase: A rapid assay for telomerase activity on telomeres",
abstract = "Most immortal cell lines derived from human cancers or transformed in vitro maintain telomeres by endogenous expression of telomerase. In the present work, immortal cells that already express endogenous telomerase activity were induced to overexpress an exogenous telomerase (hTERT) and were analyzed for changes in telomere length. Introduction of hTERT into telomerase-positive immortal cell lines results in elevated telomerase activity as measured by the TRAP assay, leading to elongated telomeres in the cell lines tested. We explore possibilities for regulatory differences among the cell lines, including the level of over-expression of the catalytic subunit hTERT and the endogenous levels of telomere binding proteins. Reducing levels of hTERT expression with a construct containing an inefficient translation initiation sequence provided sufficient telomerase expression for maximal rates of telomere elongation. Overexpression of the hTERT alters the telomere length normally maintained in these cells and provides a very useful assay for the rapid analysis of telomerase activity on its native substrate, telomeres. (C) 2000 Academic Press.",
keywords = "hTERT, hTR, Telomerase, Telomere, TRAP",
author = "McChesney, {Patricia A.} and Aisner, {Dara L.} and Frank, {Bryan C.} and Wright, {Woodring E.} and Shay, {Jerry W.}",
year = "2000",
doi = "10.1006/mcbr.2000.0229",
language = "English (US)",
volume = "3",
pages = "312--318",
journal = "Molecular Cell Biology Research Communications",
issn = "1522-4724",
publisher = "Academic Press Inc.",
number = "5",

}

TY - JOUR

T1 - Telomere dynamics in cells with introduced telomerase

T2 - A rapid assay for telomerase activity on telomeres

AU - McChesney, Patricia A.

AU - Aisner, Dara L.

AU - Frank, Bryan C.

AU - Wright, Woodring E.

AU - Shay, Jerry W.

PY - 2000

Y1 - 2000

N2 - Most immortal cell lines derived from human cancers or transformed in vitro maintain telomeres by endogenous expression of telomerase. In the present work, immortal cells that already express endogenous telomerase activity were induced to overexpress an exogenous telomerase (hTERT) and were analyzed for changes in telomere length. Introduction of hTERT into telomerase-positive immortal cell lines results in elevated telomerase activity as measured by the TRAP assay, leading to elongated telomeres in the cell lines tested. We explore possibilities for regulatory differences among the cell lines, including the level of over-expression of the catalytic subunit hTERT and the endogenous levels of telomere binding proteins. Reducing levels of hTERT expression with a construct containing an inefficient translation initiation sequence provided sufficient telomerase expression for maximal rates of telomere elongation. Overexpression of the hTERT alters the telomere length normally maintained in these cells and provides a very useful assay for the rapid analysis of telomerase activity on its native substrate, telomeres. (C) 2000 Academic Press.

AB - Most immortal cell lines derived from human cancers or transformed in vitro maintain telomeres by endogenous expression of telomerase. In the present work, immortal cells that already express endogenous telomerase activity were induced to overexpress an exogenous telomerase (hTERT) and were analyzed for changes in telomere length. Introduction of hTERT into telomerase-positive immortal cell lines results in elevated telomerase activity as measured by the TRAP assay, leading to elongated telomeres in the cell lines tested. We explore possibilities for regulatory differences among the cell lines, including the level of over-expression of the catalytic subunit hTERT and the endogenous levels of telomere binding proteins. Reducing levels of hTERT expression with a construct containing an inefficient translation initiation sequence provided sufficient telomerase expression for maximal rates of telomere elongation. Overexpression of the hTERT alters the telomere length normally maintained in these cells and provides a very useful assay for the rapid analysis of telomerase activity on its native substrate, telomeres. (C) 2000 Academic Press.

KW - hTERT

KW - hTR

KW - Telomerase

KW - Telomere

KW - TRAP

UR - http://www.scopus.com/inward/record.url?scp=0033843417&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033843417&partnerID=8YFLogxK

U2 - 10.1006/mcbr.2000.0229

DO - 10.1006/mcbr.2000.0229

M3 - Article

C2 - 10964756

AN - SCOPUS:0033843417

VL - 3

SP - 312

EP - 318

JO - Molecular Cell Biology Research Communications

JF - Molecular Cell Biology Research Communications

SN - 1522-4724

IS - 5

ER -