Cell cycle regulation of the activity and subcellular localization of Plk1, a human protein kinase implicated in mitotic spindle function

J Cell Biol. 1995 Jun;129(6):1617-28. doi: 10.1083/jcb.129.6.1617.

Abstract

Correct assembly and function of the mitotic spindle during cell division is essential for the accurate partitioning of the duplicated genome to daughter cells. Protein phosphorylation has long been implicated in controlling spindle function and chromosome segregation, and genetic studies have identified several protein kinases and phosphatases that are likely to regulate these processes. In particular, mutations in the serine/threonine-specific Drosophila kinase polo, and the structurally related kinase Cdc5p of Saccharomyces cerevisae, result in abnormal mitotic and meiotic divisions. Here, we describe a detailed analysis of the cell cycle-dependent activity and subcellular localization of Plk1, a recently identified human protein kinase with extensive sequence similarity to both Drosophila polo and S. cerevisiae Cdc5p. With the aid of recombinant baculoviruses, we have established a reliable in vitro assay for Plk1 kinase activity. We show that the activity of human Plk1 is cell cycle regulated, Plk1 activity being low during interphase but high during mitosis. We further show, by immunofluorescent confocal laser scanning microscopy, that human Plk1 binds to components of the mitotic spindle at all stages of mitosis, but undergoes a striking redistribution as cells progress from metaphase to anaphase. Specifically, Plk1 associates with spindle poles up to metaphase, but relocalizes to the equatorial plane, where spindle microtubules overlap (the midzone), as cells go through anaphase. These results indicate that the association of Plk1 with the spindle is highly dynamic and that Plk1 may function at multiple stages of mitotic progression. Taken together, our data strengthen the notion that human Plk1 may represent a functional homolog of polo and Cdc5p, and they suggest that this kinase plays an important role in the dynamic function of the mitotic spindle during chromosome segregation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Baculoviridae
  • Base Sequence
  • Cell Cycle / physiology*
  • Cell Cycle Proteins / metabolism
  • Cell Line
  • DNA Primers
  • Drosophila / enzymology
  • Drosophila Proteins*
  • Guanosine Triphosphate / metabolism
  • HeLa Cells
  • Humans
  • Kinetics
  • Mitosis
  • Molecular Sequence Data
  • Molecular Weight
  • Mutagenesis, Site-Directed
  • Polo-Like Kinase 1
  • Protein Kinases / analysis
  • Protein Kinases / biosynthesis
  • Protein Kinases / metabolism*
  • Protein Serine-Threonine Kinases / metabolism
  • Proto-Oncogene Proteins
  • RNA-Binding Proteins
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae Proteins
  • Spindle Apparatus / enzymology
  • Spindle Apparatus / physiology*
  • Spindle Apparatus / ultrastructure
  • Spodoptera
  • Transfection

Substances

  • CEF1 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA Primers
  • Drosophila Proteins
  • Proto-Oncogene Proteins
  • RNA-Binding Proteins
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Guanosine Triphosphate
  • Adenosine Triphosphate
  • Protein Kinases
  • polo protein, Drosophila
  • Protein Serine-Threonine Kinases