Crosslinking of an iodo-uridine-RNA hairpin to a single site on the human U1A N-terminal RNA binding domain

W. Tom Stump, Kathleen B. Hall

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

The N-terminal RNA binding domain (RBD) of the human U1A snRNP protein binds tightly and specifically to an RNA hairpin that contains a 10-nucleotide loop. The protein is one of a class of RNA binding proteins that adopts a βαββαβ global fold, which in turn forms a four-stranded antiparallel β-sheet. This sheet forms the primary binding surface for the RNA, as shown by the crosslinking results described here, and in more detail by a recently described co-crystal of this RBD with an RNA hairpin (Oubridge C, et al., 1994, Nature 372:432-438). The RNA hairpin sequence used in the crosslinking experiments, containing 5-iodo-uridine, is a variant of the normal U1 snRNA sequence which is able to form a crosslink with the protein, in contrast to the wild-type sequence, which does not. This single uridine substitution in the 10-nucleotide loop is the site of cross-linking to one tyrosine (Tyr 13) in the β1 strand of the U1A N-terminal RBD. This same uridine is also crosslinked to a mutant Tyr 13 Phe RBD, at this Phe 13 substitution.

Original languageEnglish
Pages (from-to)55-63
Number of pages9
JournalRNA
Volume1
Issue number1
StatePublished - Dec 1 1995

Keywords

  • 5-iodo-uridine RNA
  • Protein mutagenesis
  • Site-specific crosslinking
  • U1A RBD

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