EVI1 carboxy-terminal phosphorylation is ATM-mediated and sustains transcriptional modulation and self-renewal via enhanced CtBP1 association.

Roberto Paredes,Axel Schambach,Marion Schneider,Stefan Meyer,Kathleen Connors,Harald Löffler,Joanne Muter,Stella Pearson,Olga Kustikova,Adam Stevens,Hsiang Ying Teng,Andrew Pierce,Richard D Unwin ,John Chadwick ,James R Kelly ,Daniel J White ,Andrew J.k Williamson ,Anthony D Whetton ,Simon C Lovell ,Tim C P Somervaille ,Henri J Van De Vrugt ,Hannah Smith ,Daniel H Wiseman

doi:10.1093/nar/gky536

Abstract

The transcriptional regulator EVI1 has an essential role in early hematopoiesis and development. However, aberrantly high expression of EVI1 has potent oncogenic properties and confers poor prognosis and chemo-resistance in leukemia and solid tumors. To investigate to what extent EVI1 function might be regulated by post-translational modifications we carried out mass spectrometry- and antibody-based analyses and uncovered an ATM-mediated double phosphorylation of EVI1 at the carboxy-terminal S858/S860 SQS motif. In the presence of genotoxic stress EVI1-WT (SQS), but not site mutated EVI1-AQA was able to maintain transcriptional patterns and transformation potency, while under standard conditions carboxy-terminal mutation had no effect. Maintenance of hematopoietic progenitor cell clonogenic potential was profoundly impaired with EVI1-AQA compared with EVI1-WT, in particular in the presence of genotoxic stress. Exploring mechanistic events underlying these observations, we showed that after genotoxic stress EVI1-WT, but not EVI1-AQA increased its level of association with its functionally essential interaction partner CtBP1, implying a role for ATM in regulating EVI1 protein interactions via phosphorylation. This aspect of EVI1 regulation is therapeutically relevant, as chemotherapy-induced genotoxicity might detrimentally sustain EVI1 function via stress response mediated phosphorylation, and ATM-inhibition might be of specific targeted benefit in EVI1-overexpressing malignancies.

Highlights

The GEO access number relating to the RNAseq data is incorrect
Tel: +44 161 446 3090; Fax: +44 161 446 3091; Email: stefan.meyer@manchester.ac.uk †The authors wish it to be known that, in their opinion, the first two authors should be regarded as Joint First Authors
Harald Loffler, Zentrum fur Innere Medizin III, Marienhospital, Stuttgart, Germany

Summary

Introduction

EVI1 carboxy-terminal phosphorylation is ATM-mediated and sustains transcriptional modulation and self-renewal via enhanced CtBP1 association Roberto Paredes1,2,†, Marion Schneider1,2,†, Adam Stevens2,3, Daniel J.

Results

Conclusion