Cell cycle markers for live cell analyses

Hariharan P. Easwaran, Heinrich Leonhardt, M. Cristina Cardoso

Research output: Contribution to journalArticle

Abstract

Many cellular processes are regulated by cell cycle dependent changes in protein dynamics and localization. Studying these changes in vivo requires methods to distinguish the different cell cycle stages. Here we demonstrate the use of DNA Ligase I fused to DsRed1 as an in situ marker to identify S phase and the subsequent transition to G2 in live cells. Using this marker, we observed changes in the nuclear distribution of Dnmt1 during cell cycle progression. Based on the different nuclear distribution of DNA Ligase I and Dnmt1 in G2 and G1, we demonstrate that the combination of both proteins allows the direct discrimination of all cell cycle phases using either immunostainings or fusions with fluorescent proteins. These markers are new tools to directly study cell cycle dependent processes in both, fixed and living cells.

Original languageEnglish (US)
Pages (from-to)453-455
Number of pages3
JournalCell Cycle
Volume4
Issue number3
DOIs
StatePublished - Mar 2005
Externally publishedYes

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Keywords

  • Cell cycle
  • DNA ligase I
  • DNA replication
  • Dnmt1
  • G phase marker
  • Live cell microscopy
  • PCNA
  • S phase marker

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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