title: Structural basis for dodecameric assembly states and conformational plasticity of the full-length AAA+ ATPases Rvb1 · Rvb2.
abstract: As building blocks of diverse macromolecular complexes, the AAA+ ATPases Rvb1 and Rvb2 are crucial for many cellular activities including cancer-related processes. Their oligomeric structure and function remain unclear. We report the crystal structures of full-length heteromeric Rvb1·Rvb2 complexes in distinct nucleotide binding states. Chaetomium thermophilum Rvb1·Rvb2 assemble into hexameric rings of alternating molecules and into stable dodecamers. Intriguingly, the characteristic oligonucleotide-binding (OB) fold domains (DIIs) of Rvb1 and Rvb2 occupy unequal places relative to the compact AAA+ core ring. While Rvb1's DII forms contacts between hexamers, Rvb2's DII is rotated 100° outward, occupying lateral positions. ATP was retained bound to Rvb1 but not Rvb2 throughout purification, suggesting nonconcerted ATPase activities and nucleotide binding. Significant conformational differences between nucleotide-free and ATP-/ADP-bound states in the crystal structures and in solution suggest that the functional role of Rvb1·Rvb2 is mediated by highly interconnected structural switches. Our structures provide an atomic framework for dodecameric states and Rvb1·Rvb2's conformational plasticity.
authors: Department of Biochemistry, Gene Center of the Ludwig-Maximilians University Munich, 81377 Munich, Germany.
authors: Kristina
authors: K
authors: Lakomek
authors: Department of Biochemistry, Gene Center of the Ludwig-Maximilians University Munich, 81377 Munich, Germany.
authors: Gabriele
authors: G
authors: Stoehr
authors: Department of Biochemistry, Gene Center of the Ludwig-Maximilians University Munich, 81377 Munich, Germany.
authors: Alessandro
authors: A
authors: Tosi
authors: Department of Biochemistry, Gene Center of the Ludwig-Maximilians University Munich, 81377 Munich, Germany.
authors: Monika
authors: M
authors: Schmailzl
authors: Department of Biochemistry, Gene Center of the Ludwig-Maximilians University Munich, 81377 Munich, Germany; Center for Integrated Protein Sciences, Ludwig-Maximilians University Munich, Feodor-Lynen-Str. 25, 81377 Munich, Germany. Electronic address: hopfner@genzentrum.lmu.de.
authors: Karl-Peter
authors: KP
authors: Hopfner
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry; D020134:Catalytic Domain; D002597:Chaetomium / Q000201:enzymology*; D018360:Crystallography, X-Ray; D005656:Fungal Proteins / Q000737:chemistry*; D008958:Models, Molecular; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D053838:Scattering, Small Angle
chemical_list: D005656:Fungal Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases