Altered subcellular localization of tumor-specific cyclin E isoforms affects cyclin-dependent kinase 2 complex formation and proteasomal regulation

Cancer Res. 2009 Apr 1;69(7):2817-25. doi: 10.1158/0008-5472.CAN-08-4182. Epub 2009 Mar 24.

Abstract

In tumors, alternative translation and posttranslational proteolytic cleavage of full-length cyclin E (EL) produces tumorigenic low molecular weight cyclin E (LMW-E) isoforms that lack a portion of the EL amino-terminus containing a nuclear localization sequence. Therefore, we hypothesized that LMW-E isoforms have altered subcellular localization. To explore our hypothesis, we compared EL versus LMW-E localization in cell lysates and in vivo using fractionation and protein complementation assays. Our results reveal that LMW-E isoforms preferentially accumulate in the cytoplasm where they bind the cyclin E kinase partner, cyclin-dependent kinase 2 (Cdk2), and have associated kinase activity. The nuclear ubiquitin ligase Fbw7 targets Cdk2-bound cyclin E for degradation; thus, we examined if altered subcellular localization affected LMW-E degradation. We found that cytoplasmic LMW-E/Cdk2 was less susceptible to Fbw7-mediated degradation. One implication of our findings is that altered LMW-E and LMW-E/Cdk2 subcellular localization may lead to aberrant LMW-E protein interactions, regulation, and activity, ultimately contributing to LMW-E tumorigenicity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / metabolism
  • Cell Nucleus / enzymology
  • Cell Nucleus / metabolism
  • Cyclin E / metabolism*
  • Cyclin-Dependent Kinase 2 / metabolism*
  • Cytoplasm / enzymology
  • Cytoplasm / metabolism
  • F-Box Proteins / metabolism
  • F-Box-WD Repeat-Containing Protein 7
  • Humans
  • Molecular Weight
  • Neoplasms / enzymology
  • Neoplasms / metabolism*
  • Proteasome Endopeptidase Complex / metabolism*
  • Protein Isoforms
  • Subcellular Fractions / enzymology
  • Subcellular Fractions / metabolism
  • Tumor Cells, Cultured
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • Cell Cycle Proteins
  • Cyclin E
  • F-Box Proteins
  • F-Box-WD Repeat-Containing Protein 7
  • FBXW7 protein, human
  • Protein Isoforms
  • Ubiquitin-Protein Ligases
  • CDK2 protein, human
  • Cyclin-Dependent Kinase 2
  • Proteasome Endopeptidase Complex