A novel interaction of Ecdysoneless (ECD) protein with R2TP complex component RUVBL1 is required for the functional role of ECD in cell cycle progression

Riyaz A. Mir, Aditya Bele, Sameer Mirza, Shashank Srivastava, Appolinaire A. Olou, Shalis A. Ammons, Jun Hyun Kim, Channabasavaiah B Gurumurthy, Fang Qiu, Hamid Band, Vimla Band

Research output: Contribution to journalArticle

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Abstract

Ecdysoneless (ECD) is an evolutionarily conserved protein whose germ line deletion is embryonic lethal. Deletion of Ecd in cells causes cell cycle arrest, which is rescued by exogenous ECD, demonstrating a requirement of ECD for normal mammalian cell cycle progression. However, the exact mechanism by which ECD regulates cell cycle is unknown. Here, we demonstrate that ECD protein levels and subcellular localization are invariant during cell cycle progression, suggesting a potential role of posttranslational modifications or protein-protein interactions. Since phosphorylated ECD was recently shown to interact with the PIH1D1 adaptor component of the R2TP cochaperone complex, we examined the requirement of ECD phosphorylation in cell cycle progression. Notably, phosphorylation-deficient ECD mutants that failed to bind to PIH1D1 in vitro fully retained the ability to interact with the R2TP complex and yet exhibited a reduced ability to rescue Ecd-deficient cells from cell cycle arrest. Biochemical analyses demonstrated an additional phosphorylation-independent interaction of ECD with the RUVBL1 component of the R2TP complex, and this interaction is essential for ECD's cell cycle progression function. These studies demonstrate that interaction of ECD with RUVBL1, and its CK2-mediated phosphorylation, independent of its interaction with PIH1D1, are important for its cell cycle regulatory function.

Original languageEnglish (US)
Pages (from-to)886-899
Number of pages14
JournalMolecular and cellular biology
Volume36
Issue number6
DOIs
StatePublished - Jan 1 2016

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Cell Cycle
Phosphorylation
Proteins
Cell Cycle Checkpoints
Post Translational Protein Processing
Germ Cells

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

A novel interaction of Ecdysoneless (ECD) protein with R2TP complex component RUVBL1 is required for the functional role of ECD in cell cycle progression. / Mir, Riyaz A.; Bele, Aditya; Mirza, Sameer; Srivastava, Shashank; Olou, Appolinaire A.; Ammons, Shalis A.; Kim, Jun Hyun; Gurumurthy, Channabasavaiah B; Qiu, Fang; Band, Hamid; Band, Vimla.

In: Molecular and cellular biology, Vol. 36, No. 6, 01.01.2016, p. 886-899.

Research output: Contribution to journalArticle

Mir, Riyaz A. ; Bele, Aditya ; Mirza, Sameer ; Srivastava, Shashank ; Olou, Appolinaire A. ; Ammons, Shalis A. ; Kim, Jun Hyun ; Gurumurthy, Channabasavaiah B ; Qiu, Fang ; Band, Hamid ; Band, Vimla. / A novel interaction of Ecdysoneless (ECD) protein with R2TP complex component RUVBL1 is required for the functional role of ECD in cell cycle progression. In: Molecular and cellular biology. 2016 ; Vol. 36, No. 6. pp. 886-899.
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abstract = "Ecdysoneless (ECD) is an evolutionarily conserved protein whose germ line deletion is embryonic lethal. Deletion of Ecd in cells causes cell cycle arrest, which is rescued by exogenous ECD, demonstrating a requirement of ECD for normal mammalian cell cycle progression. However, the exact mechanism by which ECD regulates cell cycle is unknown. Here, we demonstrate that ECD protein levels and subcellular localization are invariant during cell cycle progression, suggesting a potential role of posttranslational modifications or protein-protein interactions. Since phosphorylated ECD was recently shown to interact with the PIH1D1 adaptor component of the R2TP cochaperone complex, we examined the requirement of ECD phosphorylation in cell cycle progression. Notably, phosphorylation-deficient ECD mutants that failed to bind to PIH1D1 in vitro fully retained the ability to interact with the R2TP complex and yet exhibited a reduced ability to rescue Ecd-deficient cells from cell cycle arrest. Biochemical analyses demonstrated an additional phosphorylation-independent interaction of ECD with the RUVBL1 component of the R2TP complex, and this interaction is essential for ECD's cell cycle progression function. These studies demonstrate that interaction of ECD with RUVBL1, and its CK2-mediated phosphorylation, independent of its interaction with PIH1D1, are important for its cell cycle regulatory function.",
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