title: Structural and functional insights into a dodecameric molecular machine - the RuvBL1/RuvBL2 complex.
abstract: RuvBL1 (RuvB-like 1) and its homolog RuvBL2 are evolutionarily highly conserved AAA(+) ATPases essential for many cellular activities. They play an important role in chromatin remodeling, transcriptional regulation and DNA damage repair. RuvBL1 and RuvBL2 are overexpressed in different types of cancer and interact with major oncogenic factors, such as β-catenin and c-Myc regulating their function. We solved the first three-dimensional crystal structure of the human RuvBL complex with a truncated domain II and show that this complex is competent for helicase activity. The structure reveals a dodecamer consisting of two heterohexameric rings with alternating RuvBL1 and RuvBL2 monomers bound to ADP/ATP, that interact with each other via the retained part of domain II. The dodecameric quaternary structure of the R1ΔDII/R2ΔDII complex observed in the crystal structure was confirmed by small-angle X-ray scattering analysis. Interestingly, truncation of domain II led to a substantial increase in ATP consumption of RuvBL1, RuvBL2 and their complex. In addition, we present evidence that DNA unwinding of the human RuvBL proteins can be auto-inhibited by domain II, which is not present in the homologous bacterial helicase RuvB. Our data give new insights into the molecular arrangement of RuvBL1 and RuvBL2 and strongly suggest that in vivo activities of these highly interesting therapeutic drug targets are regulated by cofactors inducing conformational changes via domain II in order to modulate the enzyme complex into its active state.
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Sabine
authors.initials: S
authors.lastname: Gorynia
authors.forename: Tiago M
authors.initials: TM
authors.lastname: Bandeiras
authors.forename: Filipa G
authors.initials: FG
authors.lastname: Pinho
authors.forename: Colin E
authors.initials: CE
authors.lastname: McVey
authors.forename: Clemens
authors.initials: C
authors.lastname: Vonrhein
authors.forename: Adam
authors.initials: A
authors.lastname: Round
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Svergun
authors.forename: Peter
authors.initials: P
authors.lastname: Donner
authors.forename: Pedro M
authors.initials: PM
authors.lastname: Matias
authors.forename: Maria Arménia
authors.initials: MA
authors.lastname: Carrondo
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D002352:Carrier Proteins / Q000737:chemistry*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004265:DNA Helicases / Q000737:chemistry*; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
chemical_list: D002352:Carrier Proteins; D046911:Macromolecular Substances; D000074183:ATPases Associated with Diverse Cellular Activities; D004265:DNA Helicases; C486549:RUVBL1 protein, human; C122079:RUVBL2 protein, human