TY - JOUR
T1 - Pih1p-Tah1p Puts a Lid on Hexameric AAA+ ATPases Rvb1/2p
AU - Tian, Shaoxiong
AU - Yu, Ge
AU - He, Huan
AU - Zhao, Yu
AU - Liu, Peilu
AU - Marshall, Alan G.
AU - Demeler, Borries
AU - Stagg, Scott M.
AU - Li, Hong
N1 - Funding Information:
We thank John Spear for assistance in EM data collection, Shelby Davis for plasmid preparation, and B. Washburn and C. Pye of the Molecular Cloning Facility at FSU for the cloning experiments. This work was supported by NIH grants R01 GM66958 and R01 GM099604 to H.L. and NSF Division of Materials Research through DMR-11-57490 to A.G.M. The UltraScan development is supported by National Science Foundation grant ACI-1339649 (to B.D.), and AUC data analysis is supported by XSEDE allocation grant MCB-070039 (to B.D.).
PY - 2017/10/3
Y1 - 2017/10/3
N2 - The Saccharomyces cerevisiae (Sc) R2TP complex affords an Hsp90-mediated and nucleotide-driven chaperone activity to proteins of small ribonucleoprotein particles (snoRNPs). The current lack of structural information on the ScR2TP complex, however, prevents a mechanistic understanding of this biological process. We characterized the structure of the ScR2TP complex made up of two AAA+ ATPases, Rvb1/2p, and two Hsp90 binding proteins, Tah1p and Pih1p, and its interaction with the snoRNP protein Nop58p by a combination of analytical ultracentrifugation, isothermal titration calorimetry, chemical crosslinking, hydrogen-deuterium exchange, and cryoelectron microscopy methods. We find that Pih1p-Tah1p interacts with Rvb1/2p cooperatively through the nucleotide-sensitive domain of Rvb1/2p. Nop58p further binds Pih1p-Tahp1 on top of the dome-shaped R2TP. Consequently, nucleotide binding releases Pih1p-Tah1p from Rvb1/2p, which offers a mechanism for nucleotide-driven binding and release of snoRNP intermediates. Tian et al. report that AAA+ ATPases, Rvb1/2p, though previously shown to form both dodecamer and hexamer, form a hexameric base for binding co-chaperone Pih1p-Tah1p. Tian et al. also showed that the binding interface is made of flexible and nucleotide-sensitive insertions within Rvb1/2p, which offers the basis for nucleotide-driven client release.
AB - The Saccharomyces cerevisiae (Sc) R2TP complex affords an Hsp90-mediated and nucleotide-driven chaperone activity to proteins of small ribonucleoprotein particles (snoRNPs). The current lack of structural information on the ScR2TP complex, however, prevents a mechanistic understanding of this biological process. We characterized the structure of the ScR2TP complex made up of two AAA+ ATPases, Rvb1/2p, and two Hsp90 binding proteins, Tah1p and Pih1p, and its interaction with the snoRNP protein Nop58p by a combination of analytical ultracentrifugation, isothermal titration calorimetry, chemical crosslinking, hydrogen-deuterium exchange, and cryoelectron microscopy methods. We find that Pih1p-Tah1p interacts with Rvb1/2p cooperatively through the nucleotide-sensitive domain of Rvb1/2p. Nop58p further binds Pih1p-Tahp1 on top of the dome-shaped R2TP. Consequently, nucleotide binding releases Pih1p-Tah1p from Rvb1/2p, which offers a mechanism for nucleotide-driven binding and release of snoRNP intermediates. Tian et al. report that AAA+ ATPases, Rvb1/2p, though previously shown to form both dodecamer and hexamer, form a hexameric base for binding co-chaperone Pih1p-Tah1p. Tian et al. also showed that the binding interface is made of flexible and nucleotide-sensitive insertions within Rvb1/2p, which offers the basis for nucleotide-driven client release.
KW - AAA+ proteins
KW - FT-ICR
KW - FTMS
KW - Fourier transform mass spectrometry
KW - Rvb1
KW - Rvb2
KW - analytical ultracentrifugation
KW - cryoelectron microscopy
KW - hydrogen/deuterium exchange
KW - ion cyclotron resonance
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U2 - 10.1016/j.str.2017.08.002
DO - 10.1016/j.str.2017.08.002
M3 - Article
C2 - 28919439
AN - SCOPUS:85029437760
VL - 25
SP - 1519-1529.e4
JO - Structure with Folding & design
JF - Structure with Folding & design
SN - 0969-2126
IS - 10
ER -