Screening of cell cycle fusion proteins to identify kinase signaling networks

Michelle Trojanowsky, Dusica Vidovic, Scott Simanski, Clara Penas, Stephan C Schuerer, Nagi Ayad

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Kinase signaling networks are well-established mediators of cell cycle transitions. However, how kinases interact with the ubiquitin proteasome system (UPS) to elicit protein turnover is not fully understood. We sought a means of identifying kinase-substrate interactions to better understand signaling pathways controlling protein degradation. Our prior studies used a luciferase fusion protein to uncover kinase networks controlling protein turnover. In this study, we utilized a similar approach to identify pathways controlling the cell cycle protein p27Kip1. We generated a p27Kip1-luciferase fusion and expressed it in cells incubated with compounds from a library of pharmacologically active compounds. We then compared the relative effects of the compounds on p27Kip1-luciferase fusion stabilization. This was combined with in silico kinome profiling to identify potential kinases inhibited by each compound. This approach effectively uncovered known kinases regulating p27Kip1 turnover. Collectively, our studies suggest that this parallel screening approach is robust and can be applied to fully understand kinase-ubiquitin pathway interactions.

Original languageEnglish (US)
Pages (from-to)1274-1281
Number of pages8
JournalCell Cycle
Volume14
Issue number8
DOIs
StatePublished - Apr 15 2015

Fingerprint

Cell Cycle Proteins
Cell Fusion
Phosphotransferases
Luciferases
Ubiquitin
Proteins
Proteasome Endopeptidase Complex
Computer Simulation
Proteolysis
Cell Cycle

Keywords

  • cell cycle
  • degradation
  • kinases
  • signaling networks
  • ubiquitin

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Developmental Biology

Cite this

Screening of cell cycle fusion proteins to identify kinase signaling networks. / Trojanowsky, Michelle; Vidovic, Dusica; Simanski, Scott; Penas, Clara; Schuerer, Stephan C; Ayad, Nagi.

In: Cell Cycle, Vol. 14, No. 8, 15.04.2015, p. 1274-1281.

Research output: Contribution to journalArticle

Trojanowsky, Michelle ; Vidovic, Dusica ; Simanski, Scott ; Penas, Clara ; Schuerer, Stephan C ; Ayad, Nagi. / Screening of cell cycle fusion proteins to identify kinase signaling networks. In: Cell Cycle. 2015 ; Vol. 14, No. 8. pp. 1274-1281.
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