The DEAD box protein eIF4A. 2. A cycle of nucleotide and RNA-dependent conformational changes.
Limited proteolysis experiments have been carried out with the DEAD box protein eIF4A. The results suggest that there is a substantial conformational change in eIF4A upon binding single-stranded RNA. Binding of ADP induces conformational changes in the free enzyme and the enzyme.RNA complex, and binding of the ATP analogue AMP-PNP induces a conformational change in the enzyme.RNA complex. The presence or absence of the gamma-phosphate on the bound nucleotide acts as a switch, presumably via the Walker motifs, that mediates changes in protein conformation and, as described in the preceding paper in this issue, also mediates changes in RNA affinity. Thus, these results suggest that there is a series of changes in conformation and substrate affinity throughout the ATP hydrolysis reaction cycle. A model is proposed in which eIF4A and the eIF4A-like domains of the DEAD box proteins act as ATP-driven conformational switches or motors that produce movements or structural rearrangements of attached protein domains or associated proteins. These movements could then be used to rearrange RNA structures or RNA.protein complexes.
Lorsch
J R
JR
Department of Biochemistry, Beckman Center, B400, Stanford University, Stanford, California 94305-5307, USA.
Herschlag
D
D
D000227:Adenine Nucleotides / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002021:Buffers; D039601:Eukaryotic Initiation Factor-4A; D006868:Hydrolysis; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D012313:RNA / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics; D014357:Trypsin
D000227:Adenine Nucleotides; D002021:Buffers; D008024:Ligands; D010448:Peptide Initiation Factors; D011994:Recombinant Proteins; D012313:RNA; D000255:Adenosine Triphosphate; D039601:Eukaryotic Initiation Factor-4A; D014357:Trypsin; D000251:Adenosine Triphosphatases