PDBsum entry 2i7t

Hydrolase, RNA binding protein

PDB id: 2i7t

Contents

Protein chain

404 a.a. *

Ligands

SO4

Metals

_ZN ×2

Waters ×161

* Residue conservation analysis

PDB id:

2i7t

Name:

Hydrolase, RNA binding protein

Title:

Structure of human cpsf-73

Structure:

Cleavage and polyadenylation specificity factor 73 kda subunit. Chain: a. Synonym: cpsf 73 kda subunit. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: cpsf3, cpsf73. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.10Å R-factor: 0.231 R-free: 0.276

Authors:

C.R.Mandel,H.Zhang,L.Tong

Key ref:

C.R.Mandel et al. (2006). Polyadenylation factor CPSF-73 is the pre-mRNA 3'-end-processing endonuclease. Nature, 444, 953-956. PubMed id: 17128255 DOI: 10.1038/nature05363

Date:

31-Aug-06 Release date: 30-Jan-07

Protein chain

Q9UKF6  (CPSF3_HUMAN) -  Cleavage and polyadenylation specificity factor subunit 3 from Homo sapiens

Seq: 684 a.a.

Struc: 404 a.a.

Enzyme reactions

• Enzyme class: E.C.3.1.27.- - ?????

DOI no: 10.1038/nature05363

Nature 444:953-956 (2006)

PubMed id: 17128255

Polyadenylation factor CPSF-73 is the pre-mRNA 3'-end-processing endonuclease.

C.R.Mandel, S.Kaneko, H.Zhang, D.Gebauer, V.Vethantham, J.L.Manley, L.Tong.

ABSTRACT

Most eukaryotic messenger RNA precursors (pre-mRNAs) undergo extensive maturational processing, including cleavage and polyadenylation at the 3'-end. Despite the characterization of many proteins that are required for the cleavage reaction, the identity of the endonuclease is not known. Recent analyses indicated that the 73-kDa subunit of cleavage and polyadenylation specificity factor (CPSF-73) might be the endonuclease for this and related reactions, although no direct data confirmed this. Here we report the crystal structures of human CPSF-73 at 2.1 A resolution, complexed with zinc ions and a sulphate that might mimic the phosphate group of the substrate, and the related yeast protein CPSF-100 (Ydh1) at 2.5 A resolution. Both CPSF-73 and CPSF-100 contain two domains, a metallo-beta-lactamase domain and a novel beta-CASP (named for metallo-beta-lactamase, CPSF, Artemis, Snm1, Pso2) domain. The active site of CPSF-73, with two zinc ions, is located at the interface of the two domains. Purified recombinant CPSF-73 possesses RNA endonuclease activity, and mutations that disrupt zinc binding in the active site abolish this activity. Our studies provide the first direct experimental evidence that CPSF-73 is the pre-mRNA 3'-end-processing endonuclease.

Selected figure(s)

Figure 1.
Figure 1: Structures of human CPSF-73 and yeast CPSF-100 (Ydh1). a, Schematic representation of the structure of human CPSF-73. The -strands and -helices are labelled, and the two zinc atoms in the active site are shown as grey spheres. The sulphate ion is shown as a stick model. b, Schematic representation of the structure of yeast CPSF-100. The zinc atoms in the CPSF-73 structure are shown for reference. See Supplementary Fig. 2 For Schematic drawings of the metallo- -lactamase domains of the two proteins.

Figure 2.
Figure 2: The beta--CASP domain of CPSF-73 and CPSF-100. Schematic drawings of the -CASP domains of human CPSF-73 (a) and yeast CPSF-100 (b).

The above figures are reprinted by permission from Macmillan Publishers Ltd: Nature (2006, 444, 953-956) copyright 2006.

Figures were selected by an automated process.