The PET and LIM1-2 domains of testin contribute to intramolecular and homodimeric interactions

PLoS One. 2017 May 18;12(5):e0177879. doi: 10.1371/journal.pone.0177879. eCollection 2017.

Abstract

The focal adhesion protein testin is a modular scaffold and tumour suppressor that consists of an N-terminal cysteine rich (CR) domain, a PET domain of unknown function and three C-terminal LIM domains. Testin has been proposed to have an open and a closed conformation based on the observation that its N-terminal half and C-terminal half directly interact. Here we extend the testin conformational model by demonstrating that testin can also form an antiparallel homodimer. In support of this extended model we determined that the testin region (amino acids 52-233) harbouring the PET domain interacts with the C-terminal LIM1-2 domains in vitro and in cells, and assign a critical role to tyrosine 288 in this interaction.

MeSH terms

  • Amino Acid Sequence
  • Cytoskeletal Proteins / chemistry*
  • Cytoskeletal Proteins / metabolism
  • Humans
  • LIM Domain Proteins / chemistry*
  • LIM Domain Proteins / metabolism
  • Protein Domains
  • Protein Multimerization*
  • RNA-Binding Proteins

Substances

  • Cytoskeletal Proteins
  • LIM Domain Proteins
  • RNA-Binding Proteins
  • TES protein, human

Grants and funding

This work was supported by PhD scholarship to SS - Fonds Wetenschappelijk Onderzoek Vlaanderen (http://www.fwo.be/); Grant FWO-Inter UGent-UL G.0353.11N to CA (http://www.fwo.be/); PhD scholarship to EH - Fonds National de la Recherche, Luxembourg (http://fnr.lu/); and FWO-Inter UGent-UL INTER/FWO/10/01 to Evelyne Friederich, taken over by ESR (http://fnr.lu/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.