TY - JOUR
T1 - The Hsp104 N-Terminal Domain Enables Disaggregase Plasticity and Potentiation
AU - Sweeny, Elizabeth A.
AU - Jackrel, Meredith E.
AU - Go, Michelle S.
AU - Sochor, Matthew A.
AU - Razzo, Beatrice M.
AU - DeSantis, Morgan E.
AU - Gupta, Kushol
AU - Shorter, James
N1 - Funding Information:
We thank Sue Lindquist, Walid Houry, Aaron Gitler, Martin Duennwald, and Laura Castellano for kindly providing reagents; Marta Carroni and Helen Saibil for sharing HAP model coordinates; Greg Van Duyne for help with SAXS; and Hiro Tsuruta, Lin Yang, and Marc Allaire for beamline assistance. This work was funded by American Heart Association predoctoral (E.A.S.) and postdoctoral (M.E.J.) fellowships; NIH grants T32GM008275 (E.A.S. and M.A.S.), T32GM071339 (M.E.D.), F31NS079009 (M.E.D.), DP2OD002177 (J.S.), and R01GM099836 (J.S.); an Ellison Medical Foundation New Scholar in Aging Award, Target ALS, Muscular Dystrophy Association (MDA277268), and The Robert Packard Center for ALS Research at Johns Hopkins University (J.S.).
Publisher Copyright:
© 2015 Elsevier Inc.
PY - 2015/3/5
Y1 - 2015/3/5
N2 - The structural basis by which Hsp104 dissolves disordered aggregates and prions is unknown. A single subunit within the Hsp104 hexamer can solubilize disordered aggregates, whereas prion dissolution requires collaboration by multiple Hsp104 subunits.Here, we establish that the poorly understood Hsp104 N-terminal domain (NTD) enables this operational plasticity. Hsp104 lacking the NTD (Hsp104δN) dissolves disordered aggregates but cannot dissolve prions or be potentiated by activating mutations. We define how Hsp104δN invariably stimulates Sup35 prionogenesis by fragmenting prions without solubilizing Sup35, whereas Hsp104 couples Sup35 prion fragmentation and dissolution. Volumetric reconstruction of Hsp104 hexamers in ATPγS, ADP-AlFx (hydrolysis transition state mimic), and ADP via small-angle X-ray scattering revealed a peristaltic pumping motion upon ATP hydrolysis, which drives directional substrate translocation through the central Hsp104 channel and is profoundly altered in Hsp104δN. We establish that the Hsp104 NTD enables cooperative substrate translocation, which is critical for prion dissolution and potentiated disaggregase activity.
AB - The structural basis by which Hsp104 dissolves disordered aggregates and prions is unknown. A single subunit within the Hsp104 hexamer can solubilize disordered aggregates, whereas prion dissolution requires collaboration by multiple Hsp104 subunits.Here, we establish that the poorly understood Hsp104 N-terminal domain (NTD) enables this operational plasticity. Hsp104 lacking the NTD (Hsp104δN) dissolves disordered aggregates but cannot dissolve prions or be potentiated by activating mutations. We define how Hsp104δN invariably stimulates Sup35 prionogenesis by fragmenting prions without solubilizing Sup35, whereas Hsp104 couples Sup35 prion fragmentation and dissolution. Volumetric reconstruction of Hsp104 hexamers in ATPγS, ADP-AlFx (hydrolysis transition state mimic), and ADP via small-angle X-ray scattering revealed a peristaltic pumping motion upon ATP hydrolysis, which drives directional substrate translocation through the central Hsp104 channel and is profoundly altered in Hsp104δN. We establish that the Hsp104 NTD enables cooperative substrate translocation, which is critical for prion dissolution and potentiated disaggregase activity.
UR - http://www.scopus.com/inward/record.url?scp=84924037150&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2014.12.021
DO - 10.1016/j.molcel.2014.12.021
M3 - Article
C2 - 25620563
AN - SCOPUS:84924037150
SN - 1097-2765
VL - 57
SP - 836
EP - 849
JO - Molecular cell
JF - Molecular cell
IS - 5
ER -