title: MYC activity is negatively regulated by a C-terminal lysine cluster.
abstract: The MYC oncogene is not only deregulated in cancer through abnormally high levels of expression, but also through oncogenic lesions in upstream signalling cascades. Modelling MYC deregulation using signalling mutants is a productive research strategy. For example, the MYC threonine-58 to alanine substitution mutant (T58A) within MYC-homology box 1 is more transforming than wild-type (WT) MYC, because of decreased apoptosis and increased protein stability. Understanding the regulatory mechanisms controlling T58 phosphorylation has led to new approaches for the development of MYC inhibitors. In this manuscript, we have extensively characterized a MYC signalling mutant in which six lysine residues near the highly conserved MYC homology box IV and basic region have been substituted to arginines (6KR). Previous literature suggests these lysines can undergo both ubiquitylation and acetylation. We show MYC 6KR is able to fully rescue the slow growth phenotype of HO15.19 MYC-null fibroblasts, and promote cell cycle entry of serum-starved MCF10A cells. Remarkably, 6KR increased anchorage-independent colony growth compared with WT MYC in both SH-EP and MCF10A cells. Moreover, it was also more potent in promoting xenograft tumour growth of Rat1A and SH-EP cells. Combined, our data identify this region and these six lysines as important residues for the negative regulation of MYC-induced transformation. Mechanistically, we demonstrate that, unlike T58A, the increased transformation is not a result of increased protein stability or a reduced capacity for 6KR to induce apoptosis. Through expression analysis and luciferase reporter assays, we show that 6KR has increased transcriptional activity compared with WT MYC. Combined, through a comprehensive evaluation across multiple cell types, we identify an important regulatory region within MYC. A better understanding of the full scope of signalling through these residues will provide further insights into the mechanisms contributing to MYC-induced tumorigenesis and may unveil novel therapeutic strategies to target Myc in cancer.
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: A R
authors: AR
authors: Wasylishen
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: M
authors: M
authors: Kalkat
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: S S
authors: SS
authors: Kim
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: A
authors: A
authors: Pandyra
authors: Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: P-K
authors: PK
authors: Chan
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: S
authors: S
authors: Oliveri
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: E
authors: E
authors: Sedivy
authors: Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: D
authors: D
authors: Konforte
authors: Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: C
authors: C
authors: Bros
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: B
authors: B
authors: Raught
authors: 1] Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada [2] Ontario Cancer Institute, Campbell Family Institute for Cancer Research, Princess Margaret Hospital, Toronto, Ontario, Canada.
authors: L Z
authors: LZ
authors: Penn
mesh_terms: D000818:Animals; D002455:Cell Division; D005786:Gene Expression Regulation; D016259:Genes, myc; D064593:Heterografts; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D009369:Neoplasms / Q000473:pathology; D010766:Phosphorylation; D055550:Protein Stability; D051381:Rats
chemical_list: D008239:Lysine