Structural modularity of the XIST ribonucleoprotein complex

Nat Commun. 2020 Dec 2;11(1):6163. doi: 10.1038/s41467-020-20040-3.

Abstract

Long noncoding RNAs are thought to regulate gene expression by organizing protein complexes through unclear mechanisms. XIST controls the inactivation of an entire X chromosome in female placental mammals. Here we develop and integrate several orthogonal structure-interaction methods to demonstrate that XIST RNA-protein complex folds into an evolutionarily conserved modular architecture. Chimeric RNAs and clustered protein binding in fRIP and eCLIP experiments align with long-range RNA secondary structure, revealing discrete XIST domains that interact with distinct sets of effector proteins. CRISPR-Cas9-mediated permutation of the Xist A-repeat location shows that A-repeat serves as a nucleation center for multiple Xist-associated proteins and m6A modification. Thus modular architecture plays an essential role, in addition to sequence motifs, in determining the specificity of RBP binding and m6A modification. Together, this work builds a comprehensive structure-function model for the XIST RNA-protein complex, and suggests a general strategy for mechanistic studies of large ribonucleoprotein assemblies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / metabolism
  • Animals
  • Base Sequence
  • CRISPR-Cas Systems
  • Cell Line
  • Conserved Sequence
  • Cross-Linking Reagents
  • Female
  • Ficusin / chemistry
  • Formaldehyde / chemistry
  • Gene Knock-In Techniques
  • Humans
  • K562 Cells
  • Male
  • Mice
  • Mouse Embryonic Stem Cells / cytology
  • Mouse Embryonic Stem Cells / metabolism*
  • Nucleic Acid Conformation
  • Pregnancy
  • RNA, Long Noncoding / chemistry*
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism
  • Ribonucleoproteins / chemistry*
  • Ribonucleoproteins / genetics
  • Ribonucleoproteins / metabolism
  • Sequence Analysis, RNA

Substances

  • Cross-Linking Reagents
  • RNA, Long Noncoding
  • Ribonucleoproteins
  • XIST non-coding RNA
  • Formaldehyde
  • Adenine
  • Ficusin
  • 6-methyladenine