Genome-wide identification of Wig-1 mRNA targets by RIP-Seq analysis

Oncotarget. 2016 Jan 12;7(2):1895-911. doi: 10.18632/oncotarget.6557.

Abstract

RNA-binding proteins (RBPs) play important roles in the regulation of gene expression through a variety of post-transcriptional mechanisms. The p53-induced RBP Wig-1 (Zmat3) binds RNA through its zinc finger domains and enhances stability of p53 and N-Myc mRNAs and decreases stability of FAS mRNA. To identify novel Wig-1-bound RNAs, we performed RNA-immunoprecipitation followed by high-throughput sequencing (RIP-Seq) in HCT116 and Saos-2 cells. We identified 286 Wig-1-bound mRNAs common between the two cell lines. Sequence analysis revealed that AU-rich elements (AREs) are highly enriched in the 3'UTR of these Wig-1-bound mRNAs. Network enrichment analysis showed that Wig-1 preferentially binds mRNAs involved in cell cycle regulation. Moreover, we identified a 2D Wig-1 binding motif in HIF1A mRNA. Our findings confirm that Wig-1 is an ARE-BP that regulates cell cycle-related processes and provide a novel view of how Wig-1 may bind mRNA through a putative structural motif. We also significantly extend the repertoire of Wig-1 target mRNAs. Since Wig-1 is a transcriptional target of the tumor suppressor p53, these results have implications for our understanding of p53-dependent stress responses and tumor suppression.

Keywords: AREs; RIP-Seq; Wig-1; cell cycle; p53.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Bone Neoplasms / genetics*
  • Bone Neoplasms / pathology
  • Carrier Proteins / genetics*
  • Gene Ontology
  • Gene Regulatory Networks
  • HCT116 Cells
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Immunoprecipitation
  • Nuclear Proteins / genetics*
  • Osteosarcoma / genetics*
  • Osteosarcoma / pathology
  • RNA, Messenger / genetics*
  • RNA-Binding Proteins
  • Real-Time Polymerase Chain Reaction
  • Response Elements / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcriptome*
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Carrier Proteins
  • Nuclear Proteins
  • RNA, Messenger
  • RNA-Binding Proteins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • ZMAT3 protein, human