The site of 3' end formation of histone messenger RNA is a fixed distance from the downstream element recognized by the U7 snRNP

EMBO J. 1994 May 15;13(10):2432-40. doi: 10.1002/j.1460-2075.1994.tb06528.x.

Abstract

Two conserved elements direct the 3' end processing of histone messenger RNA: a stem-loop structure immediately upstream of the site of cleavage and the histone downstream element (HDE), located 12-19 nucleotides downstream of the stem-loop in the premessenger RNA. We studied the role of these two elements by systematically inserting up to 10 C residues between them in the mouse H2A-614 histone pre-mRNA. 3' End mapping of RNAs processed in vitro demonstrated that as the HDE is move downstream, the site of cleavage correspondingly moves 3'. In addition, the efficiency of processing declines. In the wild-type substrate, cleavage occurs 3' of an A residue; modest increases in the efficiency of processing of the insertion mutants were observed when an A residue was placed at the new cleavage site. The results of psoralen cross-linking studies and immunoprecipitations using anti-trimethylguanosine antibodies indicated that the decreased processing efficiency of the insertion mutants is not due to impaired binding of the U7 small nuclear ribonucleoprotein (snRNP). We conclude that the mammalian U7 snRNP acts as a molecular ruler, targeting enzymatic components of cleave histone pre-mRNAs a fixed distance from its binding site, the HDE.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Nucleus / metabolism
  • Cell-Free System
  • Cross-Linking Reagents
  • Furocoumarins
  • Guanosine / analogs & derivatives
  • Guanosine / immunology
  • HeLa Cells
  • Histones / genetics*
  • Humans
  • Mice
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Protein Binding
  • RNA Processing, Post-Transcriptional*
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / genetics
  • Regulatory Sequences, Nucleic Acid*
  • Ribonuclease H / metabolism
  • Ribonucleoproteins, Small Nuclear / metabolism*
  • Structure-Activity Relationship
  • Substrate Specificity

Substances

  • Cross-Linking Reagents
  • Furocoumarins
  • Histones
  • RNA, Messenger
  • Ribonucleoproteins, Small Nuclear
  • Guanosine
  • N(2),N(2),7-trimethylguanosine
  • Ribonuclease H