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Regulation of Cellular Signaling by CUL5 is Dependent on Its Neddylation Status

Skylar Sundquist1, Sieun Ruth Lee1, Alyssa Johnson1

  • 1Hope College, Biology and Chemistry, Paul Schaap Center, 35 E. 12th St, Holland, MI, USA.

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
|March 9, 2026
PubMed
Abstract

Insights

Neddylation of CUL5, a key protein in cell division, impacts its function. Modifying CUL5 at multiple sites influences its role in cell signaling and proliferation, offering potential therapeutic targets.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • CUL5 is a scaffold protein in ubiquitin-dependent degradation, regulating cell proliferation.
  • CUL5's function is modulated by NEDD8 modification (neddylation), a potential therapeutic target.
  • Understanding CUL5 neddylation is crucial for cancer research and therapeutic development.

Purpose of the Study:

  • To investigate the structure-function relationship of CUL5.
  • To explore how CUL5 neddylation status affects cellular processes.
  • To elucidate the role of specific lysine residues in CUL5 neddylation and function.

Main Methods:

  • Mutagenesis of CUL5 at putative neddylation sites (K724, K727, K728) and a phosphorylation site (S730).
  • Expression of wild-type and mutant CUL5 in rat endothelial cells, T47D cancer cells, and COS-1 cells.
  • Analysis of cellular proliferation, MAPK phosphorylation, ERα expression, and CUL5 neddylation, including treatment with MLN4924.

Main Results:

  • Wild-type CUL5 expression attenuated cell growth, while K724R and K724R/S730A mutants promoted growth.
  • The K724R/K727R/K728R mutant showed no significant effect on proliferation.
  • MAPK phosphorylation and ERα expression correlated with CUL5 neddylation status in T47D cells.

Conclusions:

  • NEDD8 modification of CUL5 can occur at multiple lysine residues.
  • Multi-site neddylation of CUL5 contributes to its diverse regulatory effects on cellular signaling and proliferation.
  • These findings highlight the therapeutic potential of targeting CUL5 neddylation in diseases like cancer.
Keywords:
Cul5 ; Cellular growth ; Protein modification ; Neddylation

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