Time-lapse imaging reveals dynamic relocalization of PP1gamma throughout the mammalian cell cycle.

L Trinkle-Mulcahy, PD Andrews, S Wickramasinghe, Judith Elizabeth Sleeman, AR Prescott, Yw Lam, CE Lyon, JR Swedlow, AI Lamond

Research output: Contribution to journalArticlepeer-review

Abstract

Protein phosphatase 1 (PP1) is a ubiquitous serine/threonine phosphatase that regulates many cellular processes, including cell division. When transiently expressed as fluorescent protein (FP) fusions, the three PP1 isoforms, a, beta/delta, and gamma1, are active phosphatases with distinct localization patterns. We report here the establishment and characterization of HeLa cell lines stably expressing either FP-PP1gamma or FP alone. Time-lapse imaging reveals dynamic targeting of FP-PP1gamma to specific sites throughout the cell cycle, contrasting with the diffuse pattern observed for FP alone. FP-PP1gamma shows a nucleolar accumulation during interphase. On entry into mitosis, it localizes initially at kinetochores, where it exchanges rapidly with the diffuse cytoplasmic pool. A dramatic relocalization of PP1 to the chromosome-containing regions occurs at the transition from early to late anaphase, and by telophase FP-PP1gamma also accumulates at the cleavage furrow and midbody. The changing spatio-temporal distribution of My revealed using the stable PP1 cell lines implicates it in multiple processes, including nucleolar function, the regulation of chromosome segregation and cytokinesis.

Original languageEnglish
Pages (from-to)107-17
Number of pages11
JournalMolecular Biology of the Cell
Volume14
Issue number1
DOIs
Publication statusPublished - Jan 2003

Keywords

  • PROTEIN PHOSPHATASE TYPE-1
  • SACCHAROMYCES-CEREVISIAE
  • COILED BODIES
  • HISTONE H3
  • LOCALIZATION
  • PHOSPHORYLATION
  • MITOSIS
  • KINASE
  • IDENTIFICATION
  • KINETOCHORES

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