Research Papers:

Calcium-dependent binding of Myc to calmodulin

Philipp Raffeiner _, Andrea Schraffl, Thomas Schwarz, Ruth Röck, Karin Ledolter, Markus Hartl, Robert Konrat, Eduard Stefan, Klaus Bister

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Philipp Raffeiner1,3, Andrea Schraffl1, Thomas Schwarz2, Ruth Röck1, Karin Ledolter2, Markus Hartl1, Robert Konrat2, Eduard Stefan1, Klaus Bister1

1Institute of Biochemistry and Center for Molecular Biosciences, University of Innsbruck, A-6020 Innsbruck, Austria

2Max F. Perutz Laboratories, Department of Structural and Computational Biology, University of Vienna, A-1030 Vienna, Austria

3Present address: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA

Correspondence to:

Eduard Stefan, email: eduard.stefan@uibk.ac.at

Klaus Bister, email: klaus.bister@uibk.ac.at

Keywords: oncogene, transcription factor, signal transduction, protein-protein interaction, second messenger signaling

Received: August 08, 2016     Accepted: November 21, 2016     Published: December 01, 2016


The bHLH-LZ (basic region/helix-loop-helix/leucine zipper) oncoprotein Myc and the bHLH-LZ protein Max form a binary transcription factor complex controlling fundamental cellular processes. Deregulated Myc expression leads to neoplastic transformation and is a hallmark of most human cancers. The dynamics of Myc transcription factor activity are post-translationally coordinated by defined protein-protein interactions. Here, we present evidence for a second messenger controlled physical interaction between the Ca2+ sensor calmodulin (CaM) and all Myc variants (v-Myc, c-Myc, N-Myc, and L-Myc). The predominantly cytoplasmic Myc:CaM interaction is Ca2+-dependent, and the binding site maps to the conserved bHLH domain of Myc. Ca2+-loaded CaM binds the monomeric and intrinsically disordered Myc protein with high affinity, whereas Myc:Max heterodimers show less, and Max homodimers no affinity for CaM. NMR spectroscopic analyses using alternating mixtures of 15N-labeled and unlabeled preparations of CaM and a monomeric Myc fragment containing the bHLH-LZ domain corroborate the biochemical results on the Myc:CaM interaction and confirm the interaction site mapping. In electrophoretic mobility shift assays, addition of CaM does not affect high-affinity DNA-binding of Myc:Max heterodimers. However, cell-based reporter analyses and cell transformation assays suggest that increasing CaM levels enhance Myc transcriptional and oncogenic activities. Our results point to a possible involvement of Ca2+ sensing CaM in the fine-tuning of Myc function.

Author Information

Philipp Raffeiner
Primary Contact  _

Andrea Schraffl

Thomas Schwarz

Ruth Röck

Karin Ledolter

Markus Hartl

Robert Konrat

Eduard Stefan

Klaus Bister

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