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Article Dans Une Revue EMBO Journal Année : 2004

A new a-helical extension promotes RNA binding by the dsRBD of Rnt1p RNAse III

Nicolas Leulliot
  • Fonction : Auteur
Sophie Quevillon-Cheruel
Herman van Tilbeurgh
  • Fonction : Auteur
Thomas C Leeper
  • Fonction : Auteur
Katherine S Godin
  • Fonction : Auteur
Thomas E Edwards
  • Fonction : Auteur
Snorri Tl Sigurdsson
  • Fonction : Auteur
Natasha Rozenkrants
  • Fonction : Auteur
Roland J Nagel
  • Fonction : Auteur
Manuel Ares
  • Fonction : Auteur
Gabriele Varani
  • Fonction : Auteur
  • PersonId : 1106365

Résumé

Rnt1 endoribonuclease, the yeast homolog of RNAse III, plays an important role in the maturation of a diverse set of RNAs. The enzymatic activity requires a conserved catalytic domain, while RNA binding requires the doublestranded RNA-binding domain (dsRBD) at the C-terminus of the protein. While bacterial RNAse III enzymes cleave double-stranded RNA, Rnt1p specifically cleaves RNAs that possess short irregular stem-loops containing 12-14 base pairs interrupted by internal loops and bulges and capped by conserved AGNN tetraloops. Consistent with this substrate specificity, the isolated Rnt1p dsRBD and the 30-40 amino acids that follow bind to AGNN-containing stemloops preferentially in vitro. In order to understand how Rnt1p recognizes its cognate processing sites, we have defined its minimal RNA-binding domain and determined its structure by solution NMR spectroscopy and X-ray crystallography. We observe a new carboxy-terminal helix following a canonical dsRBD structure. Removal of this helix reduces binding to Rnt1p substrates. The results suggest that this helix allows the Rnt1p dsRBD to bind to short RNA stem-loops by modulating the conformation of helix a1, a key RNA-recognition element of the dsRBD.

Dates et versions

hal-03299355 , version 1 (27-07-2021)

Identifiants

Citer

Nicolas Leulliot, Sophie Quevillon-Cheruel, Marc Graille, Herman van Tilbeurgh, Thomas C Leeper, et al.. A new a-helical extension promotes RNA binding by the dsRBD of Rnt1p RNAse III. EMBO Journal, 2004, 23 (13), pp.2468-2477. ⟨10.1038/sj.emboj.7600260⟩. ⟨hal-03299355⟩
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