ClpX(P) Generates Mechanical Force to Unfold and Translocate Its Protein Substrates

Maillard, Rodrigo A.; Chistol, Gheorghe; Sen, Maya; Righini, Maurizio; Tan, Jiongyi; Kaiser, Christian; Hodges, H. Courtney; Martin, Andreas; Bustamante, Carlos

doi:10.1016/j.cell.2011.04.010

Cited by 272 publications

(399 citation statements)

References 38 publications

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“…The former is dramatically illustrated by the forces generated by the bacterial unfoldase ClpX while engaging a polypeptide chain (Maillard et al 2011) and the latter by SNARE disassembly (Ogura and Wilkinson 2001). In our case of pontin and reptin in H/ACA RNP biogenesis, either model is conceivable (Fig.…”

Section: Discussionmentioning

confidence: 89%

Mechanism of the AAA+ ATPases pontin and reptin in the biogenesis of H/ACA RNPs

Machado-Pinilla

Liger

Leulliot

et al. 2012

RNA

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The AAA+ ATPases pontin and reptin function in a staggering array of cellular processes including chromatin remodeling, transcriptional regulation, DNA damage repair, and assembly of macromolecular complexes, such as RNA polymerase II and small nucleolar (sno) RNPs. However, the molecular mechanism for all of these AAA+ ATPase associated activities is unknown. Here we document that, during the biogenesis of H/ACA RNPs (including telomerase), the assembly factor SHQ1 holds the pseudouridine synthase NAP57/dyskerin in a viselike grip, and that pontin and reptin (as components of the R2TP complex) are required to pry NAP57 from SHQ1. Significantly, the NAP57 domain captured by SHQ1 harbors most mutations underlying X-linked dyskeratosis congenita (X-DC) implicating the interface between the two proteins as a target of this bone marrow failure syndrome. Homing in on the essential first steps of H/ACA RNP biogenesis, our findings provide the first insight into the mechanism of action of pontin and reptin in the assembly of macromolecular complexes.

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Section: Discussionmentioning

confidence: 89%

Mechanism of the AAA+ ATPases pontin and reptin in the biogenesis of H/ACA RNPs

Machado-Pinilla

Liger

Leulliot

et al. 2012

RNA

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“…Recently, the role of force has become more evident in such cellular processes as transmembrane protein transport and protein degradation (21)(22)(23)(24), stressing the importance of understanding how proteins resist force and the role of the distance to the transition state. Recent studies suggest that the molten globule plays an important role in cellular processes that involve mechanical unfolding.…”

Section: Resultsmentioning

confidence: 99%

The molten globule state is unusually deformable under mechanical force

Elms

Chodera

Bustamante

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Recently, the role of force in cellular processes has become more evident, and now with advances in force spectroscopy, the response of proteins to force can be directly studied. Such studies have found that native proteins are brittle, and thus not very deformable. Here, we examine the mechanical properties of a class of intermediates referred to as the molten globule state. Using optical trap force spectroscopy, we investigated the response to force of the native and molten globule states of apomyoglobin along different pulling axes. Unlike natively folded proteins, the molten globule state of apomyoglobin is compliant (large distance to the transition state); this large compliance means that the molten globule is more deformable and the unfolding rate is more sensitive to force (the application of force or tension will have a more dramatic effect on the unfolding rate). Our studies suggest that these are general properties of molten globules and could have important implications for mechanical processes in the cell.mechanical unfolding | protein folding M echanical force plays an important role in many biological processes, and with recent advances in force spectroscopy, the mechanical properties of single proteins can now be investigated. Force spectroscopy can be used to control the folding and unfolding behavior of a single protein molecule and follow the single trajectory of the molecule, providing previously inaccessible information about the energetics, kinetics, mechanism, and mechanical sensitivity of these processes.This force spectroscopy approach allows the determination of structural and mechanical properties of both native and partially folded states along a defined order parameter, the end-to-end extension of the molecule (Δx), which may serve as a good reaction coordinate depending on how well the states are separated along the observed order parameter (1-4). One property that can be defined along this coordinate, the distance to the transition state (Δx ‡ ), largely determines the effect of force on unfolding or refolding transitions and indicates how much the state can be deformed (in terms of its end-to-end distance) without crossing the transition state. A smaller Δx ‡ indicates the rate constant is less sensitive to force, whereas a larger Δx ‡ indicates the rate constant is more sensitive to the application of force (k ∝ exp½ðFΔx ‡ Þ∕k B T). All native proteins studied to date have a small distance to the transition state (less than 2 nm) (5-10), with small deformations resulting in unfolding ("brittle" behavior). However, many proteins populate partially folded molten globular states that are compact and contain secondary structure but lack the well-packed tertiary interactions characteristic of native proteins (11,12); such species are difficult to characterize, and their structures and roles remain largely unknown. Previous work has suggested that molten globules, with this lack of well-packed tertiary interactions, may have a larger Δx ‡ and be more compliant, and thus are more deform...

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“…As the force is applied across the protein domains the activation barrier for unfolding is lowered, increasing the probability that the protein will unfold. Studies have helped identify the role of mechanical unfolding forces in protein degradation [58][59][60]. They have uncovered details of force generation in motor proteins [61,62] and the importance of plasticity of hydrogen bond networks in regulating the mechanochemistry of cell adhesion complexes [63], cell signalling (mechanosensors) [64], force generation [61,62] and cell signalling [65,66].…”

Section: Relevant Forces For Proteinsmentioning

confidence: 99%

The physics of pulling polyproteins: a review of single molecule force spectroscopy using the AFM to study protein unfolding

2016

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One of the most exciting developments in the field of biological physics in recent years is the ability to manipulate single molecules and probe their properties and function. Since its emergence over two decades ago, single molecule force spectroscopy has become a powerful tool to explore the response of biological molecules, including proteins, DNA, RNA and their complexes, to the application of an applied force. The force versus extension response of molecules can provide valuable insight into its mechanical stability, as well as details of the underlying energy landscape. In this review we will introduce the technique of single molecule force spectroscopy using the atomic force microscope (AFM), with particular focus on its application to study proteins. We will review the models which have been developed and employed to extract information from single molecule force spectroscopy experiments. Finally, we will end with a discussion of future directions in this field.

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ClpX(P) Generates Mechanical Force to Unfold and Translocate Its Protein Substrates

Cited by 272 publications

References 38 publications

Mechanism of the AAA+ ATPases pontin and reptin in the biogenesis of H/ACA RNPs

Mechanism of the AAA+ ATPases pontin and reptin in the biogenesis of H/ACA RNPs

The molten globule state is unusually deformable under mechanical force

The physics of pulling polyproteins: a review of single molecule force spectroscopy using the AFM to study protein unfolding

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