Biophysical characterization and modeling of human Ecdysoneless (ECD) protein supports a scaffolding function

Riyaz A. Mir, Jeff Lovelace, Nicholas P. Schafer, Peter D. Simone, Admir Kellezi, Carol Kolar, Gaelle Spagnol, Paul L. Sorgen, Hamid Band, Vimla Band, Gloria E.O. Borgstahl

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

The human homolog of Drosophila ecdysoneless protein (ECD) is a p53 binding protein that stabilizes and enhances p53 functions. Homozygous deletion of mouse Ecd is early embryonic lethal and Ecd deletion delays G1-S cell cycle progression. Importantly, ECD directly interacts with the Rb tumor suppressor and competes with the E2F transcription factor for binding to Rb. Further studies demonstrated ECD is overexpressed in breast and pancreatic cancers and its overexpression correlates with poor patient survival. ECD overexpression together with Ras induces cellular transformation through upregulation of autophagy. Recently we demonstrated that CK2 mediated phosphorylation of ECD and interaction with R2TP complex are important for its cell cycle regulatory function. Considering that ECD is a component of multiprotein complexes and its crystal structure is unknown, we characterized ECD structure by circular dichroism measurements and sequence analysis software. These analyses suggest that the majority of ECD is composed of a-helices. Furthermore, small angle X-ray scattering (SAXS) analysis showed that deletion fragments, ECD(1-432) and ECD(1-534), are both well-folded and reveals that the first 400 residues are globular and the next 100 residues are in an extended cylindrical structure. Taking all these results together, we speculate that ECD acts like a structural hub or scaffolding protein in its association with its protein partners. In the future, the hypothetical model presented here for ECD will need to be tested experimentally.

Original languageEnglish (US)
Pages (from-to)195-210
Number of pages16
JournalAIMS Biophysics
Volume3
Issue number1
DOIs
StatePublished - Jan 1 2016

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Keywords

  • Circular dichroism
  • Ecdysoneless
  • Molecular modeling
  • SAXS
  • Scaffold protein
  • Structural hub

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

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