Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3)

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Oct;69(Pt 10):1107-9. doi: 10.1107/S1744309113023439. Epub 2013 Sep 28.

Abstract

Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P2₁3, with unit-cell parameters a=b=c=118.5 Å, α=β=γ=90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (VM) was 6.35 Å3 Da(-1), with a solvent content of 80.62%. Initial phases were determined by molecular replacement.

Keywords: C-terminal RNA recognition motif; ETR-3; RRM-3; human ELAV-type RNA-binding protein 3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • CELF Proteins
  • Crystallization
  • Crystallography, X-Ray
  • Humans
  • Nerve Tissue Proteins / chemistry*
  • Nerve Tissue Proteins / isolation & purification*
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / isolation & purification*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification

Substances

  • CELF Proteins
  • CELF2 protein, human
  • Nerve Tissue Proteins
  • RNA-Binding Proteins
  • Recombinant Proteins