Abstract
The hexameric AAA-ATPase Drg1 is a key factor in eukaryotic ribosome biogenesis and initiates cytoplasmic maturation of the large ribosomal subunit by releasing the shuttling maturation factor Rlp24. Drg1 monomers contain two AAA-domains (D1 and D2) that act in a concerted manner. Rlp24 release is inhibited by the drug diazaborine which blocks ATP hydrolysis in D2. The mode of inhibition was unknown. Here we show the first cryo-EM structure of Drg1 revealing the inhibitory mechanism. Diazaborine forms a covalent bond to the 2'-OH of the nucleotide in D2, explaining its specificity for this site. As a consequence, the D2 domain is locked in a rigid, inactive state, stalling the whole Drg1 hexamer. Resistance mechanisms identified include abolished drug binding and altered positioning of the nucleotide. Our results suggest nucleotide-modifying compounds as potential novel inhibitors for AAA-ATPases.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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AAA Domain
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ATPases Associated with Diverse Cellular Activities / antagonists & inhibitors
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ATPases Associated with Diverse Cellular Activities / chemistry*
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ATPases Associated with Diverse Cellular Activities / genetics
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ATPases Associated with Diverse Cellular Activities / metabolism
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Adenosine Triphosphatases / antagonists & inhibitors
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Adenosine Triphosphatases / chemistry*
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism
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Binding Sites
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Boron Compounds / chemistry*
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Boron Compounds / pharmacology
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Drug Resistance / genetics
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Enzyme Activation / drug effects
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Enzyme Activation / genetics
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Mutation
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Nucleotides / chemistry
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Saccharomyces cerevisiae Proteins / antagonists & inhibitors
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
Substances
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Boron Compounds
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Nucleotides
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Saccharomyces cerevisiae Proteins
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1,2-dihydro-1-hydroxy-6-methyl-2-(propanesulfonyl)-thieno(3,2D)(1,2,3)-diazaborine
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Adenosine Triphosphatases
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AFG2 protein, S cerevisiae
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ATPases Associated with Diverse Cellular Activities