1P1T

NMR Structure of the N-terminal RRM domain of Cleavage stimulation factor 64 KDa subunit

Summary for 1P1T

• Entry DOI: 10.2210/pdb1p1t/pdb
• Descriptor: Cleavage stimulation factor, 64 kDa subunit (1 entity in total)
• Functional Keywords: rna recognition motif, c-terminal helix, n-terminal helix, rna binding protein
• Biological source: Homo sapiens (human)
• Cellular location: Nucleus: P33240
• Total number of polymer chains: 1
• Total formula weight: 11544.84

Authors

Perez-Canadillas, J.M.,Varani, G. (deposition date: 2003-04-14, release date: 2003-08-12, Last modification date: 2024-05-22)

Primary citation

Perez-Canadillas, J.M.,Varani, G.
Recognition of GU-rich polyadenylation regulatory elements by human CstF-64 protein
Embo J., 22:2821-2830, 2003

PubMed Abstract: Vertebrate polyadenylation sites are identified by the AAUAAA signal and by GU-rich sequences downstream of the cleavage site. These are recognized by a heterotrimeric protein complex (CstF) through its 64 kDa subunit (CstF-64); the strength of this interaction affects the efficiency of poly(A) site utilization. We present the structure of the RNA-binding domain of CstF-64 containing an RNA recognition motif (RRM) augmented by N- and C-terminal helices. The C-terminal helix unfolds upon RNA binding and extends into the hinge domain where interactions with factors responsible for assembly of the polyadenylation complex occur. We propose that this conformational change initiates assembly. Consecutive Us are required for a strong CstF-GU interaction and we show how UU dinucleotides are recognized. Contacts outside the UU pocket fine tune the protein-RNA interaction and provide different affinities for distinct GU-rich elements. The protein-RNA interface remains mobile, most likely a requirement to bind many GU-rich sequences and yet discriminate against other RNAs. The structural distinction between sequences that form stable and unstable complexes provides an operational distinction between weakly and strongly processed poly(A) sites.

PubMed: 12773396
DOI: 10.1093/emboj/cdg259

Experimental method

SOLUTION NMR