Protein kinase Cι promotes UBF1-ECT2 binding on ribosomal DNA to drive rRNA synthesis and transformed growth of non-small-cell lung cancer cells

Verline Justilien, Kayla C. Lewis, Kayleah M. Meneses, Lee Jamieson, Nicole R. Murray, Alan P. Fields

Research output: Contribution to journalArticlepeer-review

Abstract

Epithelial cell-transforming sequence 2 (ECT2) is a guanine nucleotide exchange factor for Rho GTPases that is overexpressed in many cancers and involved in signal transduction pathways that promote cancer cell proliferation, invasion, and tumorigenesis. Recently, we demonstrated that a significant pool of ECT2 localizes to the nucleolus of non-small-cell lung cancer (NSCLC) cells, where it binds the transcription factor upstream binding factor 1 (UBF1) on the promoter regions of ribosomal DNA (rDNA) and activates rDNA transcription, transformed cell growth, and tumor formation. Here, we investigated the mechanism by which ECT2 engages UBF1 on rDNA promoters. Results from ECT2 mutagenesis indicated that the tandem BRCT domain of ECT2 mediates binding to UBF1. Biochemical and MS-based analyses revealed that protein kinase Cι (PKCι) directly phosphorylates UBF1 at Ser-412, thereby generating a phosphopeptide-binding epitope that binds the ECT2 BRCT domain. Lentiviral shRNA knockdown and reconstitution experiments revealed that both a functional ECT2 BRCT domain and the UBF1 Ser-412 phosphorylation site are required for UBF1-mediated ECT2 recruitment to rDNA, elevated rRNA synthesis, and transformed growth. Our findings provide critical molecular insight into ECT2-mediated regulation of rDNA transcription in cancer cells and offer a rationale for therapeutic targeting of UBF1- and ECT2-stimulated rDNA transcription for the management of NSCLC.

Original languageEnglish (US)
Pages (from-to)8214-8226
Number of pages13
JournalThe Journal of biological chemistry
Volume295
Issue number24
DOIs
StatePublished - Jun 12 2020

Keywords

  • cancer biology
  • cell proliferation
  • epithelial cell transforming protein 2 (ECT2)
  • gene regulation
  • non-small cell lung cancer
  • protein kinase C ι (PKCι)
  • protein-protein interaction
  • ribosomal DNA transcription
  • ribosomal ribonucleic acid (rRNA) (ribosomal RNA)
  • ribosome biogenesis
  • tumor cell biology
  • upstream binding factor 1 (UBF1)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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