The N-terminal domains of FLASH and Lsm11 form a 2:1 heterotrimer for histone pre-mRNA 3'-end processing

PLoS One. 2017 Oct 11;12(10):e0186034. doi: 10.1371/journal.pone.0186034. eCollection 2017.

Abstract

Unlike canonical pre-mRNAs, animal replication-dependent histone pre-mRNAs lack introns and are processed at the 3'-end by a mechanism distinct from cleavage and polyadenylation. They have a 3' stem loop and histone downstream element (HDE) that are recognized by stem-loop binding protein (SLBP) and U7 snRNP, respectively. The N-terminal domain (NTD) of Lsm11, a component of U7 snRNP, interacts with FLASH NTD and these two proteins recruit the histone cleavage complex containing the CPSF-73 endonuclease for the cleavage reaction. Here, we determined crystal structures of FLASH NTD and found that it forms a coiled-coil dimer. Using solution light scattering, we characterized the stoichiometry of the FLASH NTD-Lsm11 NTD complex and found that it is a 2:1 heterotrimer, which is supported by observations from analytical ultracentrifugation and crosslinking.

MeSH terms

  • Amino Acid Sequence
  • Biophysical Phenomena
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism*
  • Chromatography, Gel
  • Crystallography, X-Ray
  • Cysteine / genetics
  • Histones / metabolism*
  • Light
  • Mutation / genetics
  • Protein Binding
  • Protein Domains
  • Protein Multimerization*
  • Protein Structure, Secondary
  • RNA Precursors / metabolism*
  • RNA Processing, Post-Transcriptional*
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / metabolism*
  • Scattering, Radiation
  • Sequence Alignment
  • Ultracentrifugation

Substances

  • Carrier Proteins
  • Histones
  • RNA Precursors
  • RNA-Binding Proteins
  • Cysteine