Structural basis of mRNA-cap recognition by Dcp1-Dcp2.
Removal of the 5' cap on mRNA by the decapping enzyme Dcp2 is a critical step in 5'-to-3' mRNA decay. Understanding the structural basis of Dcp2 activity has been a challenge because Dcp2 is dynamic and has weak affinity for the cap substrate. Here we present a 2.6-Å-resolution crystal structure of a heterotrimer of fission yeast Dcp2, its essential activator Dcp1, and the human NMD cofactor PNRC2, in complex with a tight-binding cap analog. Cap binding is accompanied by a conformational change in Dcp2, thereby forming a composite nucleotide-binding site comprising conserved residues in the catalytic and regulatory domains. Kinetic analysis of PNRC2 revealed that a conserved short linear motif enhances both substrate affinity and the catalytic step of decapping. These findings explain why Dcp2 requires a conformational change for efficient catalysis and reveals that coactivators promote RNA binding and the catalytic step of decapping, possibly through different conformational states.
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.
Jeffrey S
JS
Mugridge
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.|Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, Warsaw, Poland.
Marcin
M
Ziemniak
Centre of New Technologies, University of Warsaw, Warsaw, Poland.
Jacek
J
Jemielity
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, California, USA.
John D
JD
Gross
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D012315:RNA Caps / Q000378:metabolism*; D020871:RNA Stability; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000378:metabolism; D012568:Schizosaccharomyces / Q000737:chemistry / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism
C527678:Dcp1 protein, S pombe; C508496:Dcp2 protein, S pombe; C437718:PNRC2 protein, human; D012315:RNA Caps; D018160:Receptors, Cytoplasmic and Nuclear; D029702:Schizosaccharomyces pombe Proteins; D015534:Trans-Activators
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