Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers

Sergeeva, Oksana A.; Chen, Bin; Haase-Pettingell, Cameron; Ludtke, Steven J.; Chiu, Wah; King, Jonathan

doi:10.1074/jbc.m112.443929

Cited by 56 publications

(89 citation statements)

References 62 publications

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“…CCT5 elutes as a homo-oligomeric, double-ring, barrel-shaped complex of ϳ1 MDa. Previous studies confirm that this TRiC-like complex has two rings with eight subunits per ring (23).…”

Section: Methodssupporting

confidence: 63%

“…In the same vein, the exogenously applied bovine recombinant purified CCT1 apical domain has been shown to penetrate cell membranes and decrease mHTT-Ex1 aggregation in vivo (22). Furthermore, recently, the purified human CCT5 subunit has been shown to form a homo-oligomeric complex and suppress mHTT aggregation in vitro (23). Interestingly, the CCT5 complex forms a double-ring barrel structure composed of 16 identical CCT5 subunits (23).…”

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

“…CCT5 Complex Purification-The human CCT5 construct was isolated as described previously (23), with a few modifications (24). Briefly, BL21(DE3)RIL Escherichia coli cells expressing human CCT5 with a His tag were lysed via French press.…”

Section: Methodsmentioning

confidence: 99%

“…Interestingly, the CCT5 complex forms a double-ring barrel structure composed of 16 identical CCT5 subunits (23). This chaperonin-like homo-oligomeric complex hydrolyzes ATP and refolds both luciferase and human ␥-Dcrystallin (23). In short, the CCT5 complex looks and acts similarly to TRiC chaperonin.…”

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

“…Furthermore, recently, the purified human CCT5 subunit has been shown to form a homo-oligomeric complex and suppress mHTT aggregation in vitro (23). Interestingly, the CCT5 complex forms a double-ring barrel structure composed of 16 identical CCT5 subunits (23). This chaperonin-like homo-oligomeric complex hydrolyzes ATP and refolds both luciferase and human ␥-Dcrystallin (23).…”

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

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Structural Mechanisms of Mutant Huntingtin Aggregation Suppression by the Synthetic Chaperonin-like CCT5 Complex Explained by Cryoelectron Tomography

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Sergeeva

Isas

et al. 2015

Journal of Biological Chemistry

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Background: Huntington disease patients show an accumulation of oligomers and fibrillar species of mutant huntingtin (mHTT). Results: Cryoelectron tomography and subvolume averaging visualizes heterogeneous mHTT oligomeric species inside the chaperonin-like CCT5 cavity. Conclusion:The structural basis of mHTT aggregation inhibition by CCT5 is through capping of fibrils and encapsulation of oligomers. Significance: These structural mechanisms inspire the development of new strategies for inhibiting mHTT aggregation.

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“…CCT5 elutes as a homo-oligomeric, double-ring, barrel-shaped complex of ϳ1 MDa. Previous studies confirm that this TRiC-like complex has two rings with eight subunits per ring (23).…”

Section: Methodssupporting

confidence: 63%

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

Section: Huntington Disease a Neurodegenerative Disorder Characterizmentioning

confidence: 99%

See 3 more Smart Citations

Structural Mechanisms of Mutant Huntingtin Aggregation Suppression by the Synthetic Chaperonin-like CCT5 Complex Explained by Cryoelectron Tomography

Darrow

Sergeeva

Isas

et al. 2015

Journal of Biological Chemistry

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Genetic expansion of chaperonin‐containing TCP‐1 (CCT/TRiC) complex subunits yields testis‐specific isoforms required for spermatogenesis in planarian flatworms

Counts

Hester

Rouhana

2017

Molecular Reproduction Devel

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Chaperonin-containing Tail-less complex polypeptide 1 (CCT) is a highly conserved, hetero-oligomeric complex that ensures proper folding of actin, tubulin, and regulators of mitosis. Eight subunits (CCT1-8) make up this complex, and every subunit has a homolog expressed in the testes and somatic tissue of the planarian flatworm Schmidtea mediterranea. Gene duplications of four subunits in the genomes of S. mediterranea and other planarian flatworms created paralogs to CCT1, CCT3, CCT4, and CCT8 that are expressed exclusively in the testes. Functional analyses revealed that each CCT subunit expressed in the S. mediterranea soma is essential for homeostatic integrity and survival, whereas sperm elongation defects were observed upon knockdown of each individual testis-specific paralog (Smed-cct1B; Smed-cct3B; Smed-cct4A; and Smed-cct8B), regardless of potential redundancy with paralogs expressed in both testes and soma (Smed-cct1A; Smed-cct3A; Smed-cct4B; and Smed-cct8A). Yet, no detriment was observed in the number of adult somatic stem cells (neoblasts) that maintain differentiated tissue in planarians. Thus, expression of all eight CCT subunits is required to execute the essential functions of the CCT complex. Furthermore, expression of the somatic paralogs in planarian testes is not sufficient to complete spermatogenesis when testis-specific paralogs are knocked down, suggesting that the evolution of chaperonin subunits may drive changes in the development of sperm structure and that correct CCT subunit stoichiometry is crucial for spermiogenesis.

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Group II archaeal chaperonin recognition of partially folded human γD‐crystallin mutants

et al. 2014

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The features in partially folded intermediates that allow the group II chaperonins to distinguish partially folded from native states remain unclear. The archaeal group II chaperonin from Methanococcus Mauripaludis (Mm-Cpn) assists the in vitro refolding of the well-characterized bsheet lens protein human cD-crystallin (HcD-Crys). The domain interface and buried cores of this Greek key conformation include side chains, which might be exposed in partially folded intermediates. We sought to assess whether particular features buried in the native state, but absent from the native protein surface, might serve as recognition signals. The features tested were (a) paired aromatic side chains, (b) side chains in the interface between the duplicated domains of HcD-Crys, and (c) side chains in the buried core which result in congenital cataract when substituted. We tested the Mm-Cpn suppression of aggregation of these HcD-Crys mutants upon dilution out of denaturant. Mm-Cpn was capable of suppressing the off-pathway aggregation of the three classes of mutants indicating that the buried residues were not recognition signals. In fact, Mm-Cpn recognized the HcD-Crys mutants better than (wild-type) WT and refolded most mutant HcD-Crys to levels twice that of WT HcD-Crys. This presumably represents the increased population or longer lifetimes of the partially folded intermediates of the mutant proteins. The results suggest that MmCpn does not recognize the features of HcD-Crys tested-paired aromatics, exposed domain interface, or destabilized core-but rather recognizes other features of the partially folded b-sheet conformation that are absent or inaccessible in the native state of HcD-Crys.

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Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers

Cited by 56 publications

References 62 publications

Structural Mechanisms of Mutant Huntingtin Aggregation Suppression by the Synthetic Chaperonin-like CCT5 Complex Explained by Cryoelectron Tomography

Structural Mechanisms of Mutant Huntingtin Aggregation Suppression by the Synthetic Chaperonin-like CCT5 Complex Explained by Cryoelectron Tomography

Genetic expansion of chaperonin‐containing TCP‐1 (CCT/TRiC) complex subunits yields testis‐specific isoforms required for spermatogenesis in planarian flatworms

Group II archaeal chaperonin recognition of partially folded human γD‐crystallin mutants

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