PDBsum entry 4yh8

Splicing

PDB id: 4yh8

Contents

Protein chains

180 a.a.

55 a.a.

Metals

_ZN ×2

Waters ×208

PDB id:

4yh8

Name:

Splicing

Title:

Structure of yeast u2af complex

Structure:

Splicing factor u2af 23 kda subunit. Chain: a. Synonym: u2 auxiliary factor 23 kda subunit,u2af23,u2 snrnp auxiliary factor small subunit. Engineered: yes. Splicing factor u2af 59 kda subunit. Chain: b. Fragment: unp residues 93-161. Synonym: u2 auxiliary factor 59 kda subunit,u2af59,u2 snrnp auxiliary

Source:

Schizosaccharomyces pombe (strain 972 / atcc 24843). Fission yeast. Organism_taxid: 284812. Strain: 972 / atcc 24843. Gene: spap8a3.06. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: prp2, mis11, spbc146.07.

Resolution:

1.70Å R-factor: 0.192 R-free: 0.216

Authors:

H.Yoshida,S.Y.Park,T.Urano,E.Obayashi

Key ref:

H.Yoshida et al. (2015). A novel 3' splice site recognition by the two zinc fingers in the U2AF small subunit. Genes Dev, 29, 1649-1660. PubMed id: 26215567 DOI: 10.1101/gad.267104.115

Date:

27-Feb-15 Release date: 12-Aug-15

Protein chain

Q09176  (U2AF1_SCHPO) -  Splicing factor U2AF 23 kDa subunit from Schizosaccharomyces pombe (strain 972 / ATCC 24843)

Seq: 216 a.a.

Struc: 180 a.a.

Protein chain

P36629  (U2AF2_SCHPO) -  Splicing factor U2AF 59 kDa subunit from Schizosaccharomyces pombe (strain 972 / ATCC 24843)

Seq: 517 a.a.

Struc: 55 a.a.

Enzyme reactions

• Enzyme class: Chains A, B: E.C.?

DOI no: 10.1101/gad.267104.115

Genes Dev 29:1649-1660 (2015)

PubMed id: 26215567

A novel 3' splice site recognition by the two zinc fingers in the U2AF small subunit.

H.Yoshida, S.Y.Park, T.Oda, T.Akiyoshi, M.Sato, M.Shirouzu, K.Tsuda, K.Kuwasako, S.Unzai, Y.Muto, T.Urano, E.Obayashi.

ABSTRACT

The pre-mRNA splicing reaction of eukaryotic cells has to be carried out extremely accurately, as failure to recognize the splice sites correctly causes serious disease. The small subunit of the U2AF heterodimer is essential for the determination of 3' splice sites in pre-mRNA splicing, and several single-residue mutations of the U2AF small subunit cause severe disorders such as myelodysplastic syndromes. However, the mechanism of RNA recognition is poorly understood. Here we solved the crystal structure of the U2AF small subunit (U2AF23) from fission yeast, consisting of an RNA recognition motif (RRM) domain flanked by two conserved CCCH-type zinc fingers (ZFs). The two ZFs are positioned side by side on the β sheet of the RRM domain. Further mutational analysis revealed that the ZFs bind cooperatively to the target RNA sequence, but the RRM domain acts simply as a scaffold to organize the ZFs and does not itself contact the RNA directly. This completely novel and unexpected mode of RNA-binding mechanism by the U2AF small subunit sheds light on splicing errors caused by mutations of this highly conserved protein.