Identification of TINO: a new evolutionarily conserved BCL-2 AU-rich element RNA-binding protein

J Biol Chem. 2004 May 7;279(19):20154-66. doi: 10.1074/jbc.M314071200. Epub 2004 Feb 9.

Abstract

Modulation of mRNA stability by regulatory cis-acting AU-rich elements (AREs) and ARE-binding proteins is an important posttranscriptional mechanism of gene expression control. We previously demonstrated that the 3'-untranslated region of BCL-2 mRNA contains an ARE that accounts for rapid BCL-2 down-regulation in response to apoptotic stimuli. We also demonstrated that the BCL-2 ARE core interacts with a number of ARE-binding proteins, one of which is AU-rich factor 1/heterogeneous nuclear ribonucleoprotein D, known for its interaction with mRNA elements of others genes. In an attempt to search for other BCL-2 mRNA-binding proteins, we used the yeast RNA three-hybrid system assay and identified a novel human protein that interacts with BCL-2 ARE. We refer to it as TINO. The predicted protein sequence of TINO reveals two amino-terminal heterogeneous nuclear ribonucleoprotein K homology motifs for nucleic acid binding and a carboxyl-terminal RING domain, endowed with a putative E3 ubiquitin-protein ligase activity. In addition the novel protein is evolutionarily conserved; the two following orthologous proteins have been identified with protein-protein BLAST: posterior end mark-3 (PEM-3) of Ciona savignyi and muscle excess protein-3 (MEX-3) of Caenorhabditis elegans. Upon binding, TINO destabilizes a chimeric reporter construct containing the BCL-2 ARE sequence, revealing a negative regulatory action on BCL-2 gene expression at the posttranscriptional level.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Apoptosis
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Caenorhabditis elegans
  • Cell Line
  • Conserved Sequence
  • DNA, Complementary / metabolism
  • Databases as Topic
  • Down-Regulation
  • Evolution, Molecular
  • Gene Expression Regulation
  • Genes, bcl-2*
  • HeLa Cells
  • Heterogeneous-Nuclear Ribonucleoprotein D / metabolism
  • Heterogeneous-Nuclear Ribonucleoprotein K / metabolism
  • Humans
  • Microscopy, Fluorescence
  • Models, Genetic
  • Molecular Sequence Data
  • Plasmids / metabolism
  • Protein Binding
  • Protein Biosynthesis
  • Protein Structure, Tertiary
  • RNA Processing, Post-Transcriptional
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / chemistry*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ribonucleases / metabolism
  • Sequence Homology, Amino Acid
  • Time Factors
  • Transfection
  • Two-Hybrid System Techniques
  • Urochordata

Substances

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • DNA, Complementary
  • Heterogeneous-Nuclear Ribonucleoprotein D
  • Heterogeneous-Nuclear Ribonucleoprotein K
  • RNA, Messenger
  • RNA-Binding Proteins
  • Ribonucleases

Associated data

  • GENBANK/AF458084