Polyadenylation Factor CPSF-73 is the Pre-mRNA 3'-end-processing Endonuclease

Journal Article · Sun Jan 01 04:00:00 EST 2006 · Nature

DOI:https://doi.org/10.1038/nature05363· OSTI ID:930590

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

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 {angstrom} 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 {angstrom} resolution. Both CPSF-73 and CPSF-100 contain two domains, a metallo-{beta}-lactamase domain and a novel -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.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 930590
• Report Number(s): BNL--80847-2008-JA
• Journal Information: Nature, Journal Name: Nature Vol. 444; ISSN 0028-0836
• Country of Publication: United States
• Language: English

Related Subjects

36 MATERIALS SCIENCE
CLEAVAGE
CRYSTAL STRUCTURE
ENDONUCLEASES
MESSENGER-RNA
MUTATIONS
PHOSPHATES
PROCESSING
PROTEINS
RESOLUTION
RNA
SPECIFICITY
YEASTS
ZINC
ZINC IONS
national synchrotron light source