Conformational changes in the GTPase modules of the signal reception particle and its receptor drive initiation of protein translocation.
During cotranslational protein targeting, two guanosine triphosphatase (GTPase) in the signal recognition particle (SRP) and its receptor (SR) form a unique complex in which hydrolyses of both guanosine triphosphates (GTP) are activated in a shared active site. It was thought that GTP hydrolysis drives the recycling of SRP and SR, but is not crucial for protein targeting. Here, we examined the translocation efficiency of mutant GTPases that block the interaction between SRP and SR at specific stages. Surprisingly, mutants that allow SRP-SR complex assembly but block GTPase activation severely compromise protein translocation. These mutations map to the highly conserved insertion box domain loops that rearrange upon complex formation to form multiple catalytic interactions with the two GTPs. Thus, although GTP hydrolysis is not required, the molecular rearrangements that lead to GTPase activation are essential for protein targeting. Most importantly, our results show that an elaborate rearrangement within the SRP-SR GTPase complex is required to drive the unloading and initiate translocation of cargo proteins.
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA. sshan@caltech.edu
Shu-ou
SO
Shan
Sowmya
S
Chandrasekar
Peter
P
Walter
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D004285:Dogs; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D011487:Protein Conformation; D021381:Protein Transport; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D018271:Signal Recognition Particle / Q000378:metabolism*
D001426:Bacterial Proteins; C087406:FtsY protein, Bacteria; D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide; D018271:Signal Recognition Particle; C036318:signal peptide receptor; D020558:GTP Phosphohydrolases