Structure of the ribosome post-recycling complex probed by chemical cross-linking and mass spectrometry.
Ribosome recycling orchestrated by the ATP binding cassette (ABC) protein ABCE1 can be considered as the final-or the first-step within the cyclic process of protein synthesis, connecting translation termination and mRNA surveillance with re-initiation. An ATP-dependent tweezer-like motion of the nucleotide-binding domains in ABCE1 transfers mechanical energy to the ribosome and tears the ribosome subunits apart. The post-recycling complex (PRC) then re-initiates mRNA translation. Here, we probed the so far unknown architecture of the 1-MDa PRC (40S/30S·ABCE1) by chemical cross-linking and mass spectrometry (XL-MS). Our study reveals ABCE1 bound to the translational factor-binding (GTPase) site with multiple cross-link contacts of the helix-loop-helix motif to the S24e ribosomal protein. Cross-linking of the FeS cluster domain to the ribosomal protein S12 substantiates an extreme lever-arm movement of the FeS cluster domain during ribosome recycling. We were thus able to reconstitute and structurally analyse a key complex in the translational cycle, resembling the link between translation initiation and ribosome recycling.
Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany.
Kristin
K
Kiosze-Becker
Structural and Computational Biology Unit, EMBL Heidelberg, Meyerhofstr. 1, 69117 Heidelberg, Germany.
Alessandro
A
Ori
Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany.
Milan
M
Gerovac
Gene Center and Center for Integrated Protein Science Munich (CiPSM), Department of Biochemistry, University of Munich, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
André
A
Heuer
Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany.
Elina
E
Nürenberg-Goloub
Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany.
Umar Jan
UJ
Rashid
Gene Center and Center for Integrated Protein Science Munich (CiPSM), Department of Biochemistry, University of Munich, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
Thomas
T
Becker
Gene Center and Center for Integrated Protein Science Munich (CiPSM), Department of Biochemistry, University of Munich, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
Roland
R
Beckmann
Structural and Computational Biology Unit, EMBL Heidelberg, Meyerhofstr. 1, 69117 Heidelberg, Germany.
Martin
0000-0002-7397-1321
M
Beck
Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany.
Robert
R
Tampé
D019843:Archaeal Proteins / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry*; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D048229:Sulfolobus solfataricus / Q000378:metabolism
D019843:Archaeal Proteins; D003432:Cross-Linking Reagents; D007506:Iron-Sulfur Proteins; D012269:Ribosomal Proteins