Biochemical and Biophysical Characterization of Recombinant Yeast Proteasome Maturation Factor Ump1

Protein degradation is essential for maintaining cellular homeostasis. The proteasome is the central enzyme responsible for non-lysosomal protein degradation in eukaryotic cells. Although proteasome assembly is not yet completely understood, a number of cofactors required for proper assembly and maturation have been identified. Ump is a short-lived maturation factor required for the efficient biogenesis of the 20S proteasome. Upon the association of the two precursor complexes, Ump is encased and is rapidly degraded after the proteolytic sites in the interior of the nascent proteasome are activated. In order to further understand the mechanisms behind proteasomal maturation, we expressed and purified yeast Ump in E. coli for biophysical and structural analysis. We show that recombinant Ump is purified as a mixture of different oligomeric species and that oligomerization is mediated by intermolecular disulfide bond formation involving the only cysteine residue present in the protein. Furthermore, a combination of bioinformatic, biochemical and structural analysis revealed that Ump shows characteristics of an intrinsically disordered protein, which might become structured only upon interaction with the proteasome subunits.

• Publication year

  2013

• Venue

  Computational and Structural Biotechnology Journal

• Publication date

  2013-04-01

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.5936/csbj.201304006PMID 24688736PMCID 3962104
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• INAUGURAL ARTICLE by a Recently Elected Academy Member:Molecular architecture of the 26S proteasome holocomplex determined by an integrative approach

  2012cited by this paper
• Near-atomic resolution structural model of the yeast 26S proteasome

  2012cited by this paper
• Complete subunit architecture of the proteasome regulatory particle

  2011cited by this paper
• Proteasome inhibitors in cancer therapy

  2011cited by this paper
• A Conserved 20S Proteasome Assembly Factor Requires a C-terminal HbYX Motif for Proteasomal Precursor Binding

  2011cited by this paper
• Multitude of binding modes attainable by intrinsically disordered proteins: a portrait gallery of disorder-based complexes.

  2011cited by this paper
• Targeting intrinsically disordered proteins in neurodegenerative and protein dysfunction diseases: another illustration of the D2 concept

  2010cited by this paper
• Chaperone-assisted assembly of the proteasome core particle.

  2010cited by this paper
• Catalytic mechanism and assembly of the proteasome.

  2009cited by this paper
• The Mysterious Unfoldome: Structureless, Underappreciated, Yet Vital Part of Any Given Proteome

  2009cited by this paper
• Recognition and processing of ubiquitin-protein conjugates by the proteasome.

  2009cited by this paper
• ALINE: a WYSIWYG protein-sequence alignment editor for publication-quality alignments.

  2009cited by this paper
• PACemakers of proteasome core particle assembly.

  2008cited by this paper
• Prediction of disordered regions in proteins based on the meta approach

  2008cited by this paper
• Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases.

  2008cited by this paper
• Dissecting β‐ring assembly pathway of the mammalian 20S proteasome

  2008cited by this paper
• The C-terminal Extension of the β7 Subunit and Activator Complexes Stabilize Nascent 20 S Proteasomes and Promote Their Maturation*

  2007cited by this paper
• The proteasome maturation protein POMP facilitates major steps of 20S proteasome formation at the endoplasmic reticulum

  2007cited by this paper
• β‐Subunit appendages promote 20S proteasome assembly by overcoming an Ump1‐dependent checkpoint

  2007cited by this paper
• 20S proteasome assembly is orchestrated by two distinct pairs of chaperones in yeast and in mammals.

  2007cited by this paper
• Possible tetramerisation of the proteasome maturation factor POMP/proteassemblin/hUmp1 and its subcellular localisation.

  2006cited by this paper
• A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes

  2005cited by this paper
• SCHIP: statistics for chromosome interphase positioning based on interchange data

  2005cited by this paper
• RONN: the bio-basis function neural network technique applied to the detection of natively disordered regions in proteins

  2005cited by this paper
• IFN-gamma-induced immune adaptation of the proteasome system is an accelerated and transient response.

  2005cited by this paper
• Assessing protein disorder and induced folding

  2005cited by this paper
• DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data

  2004cited by this paper
• Role of C-terminal Extensions of Subunits β2 and β7 in Assembly and Activity of Eukaryotic Proteasomes*

  2004cited by this paper
• Role of C-terminal extensions of subunits beta2 and beta7 in assembly and activity of eukaryotic proteasomes.

  2004cited by this paper
• Natively unfolded C‐terminal domain of caldesmon remains substantially unstructured after the effective binding to calmodulin

  2003cited by this paper
• Formation of transitory intrachain and interchain disulfide bonds accompanies the folding and oligomerization of simian virus 40 Vp1 in the cytoplasm

  2002cited by this paper
• 20 S proteasomes are imported as precursor complexes into the nucleus of yeast.

  2002cited by this paper
• The structure of the mammalian 20S proteasome at 2.75 A resolution.

  2002cited by this paper
• Denatured collapsed states in protein folding: Example of apomyoglobin

  2001cited by this paper
• Why are “natively unfolded” proteins unstructured under physiologic conditions?

  2000cited by this paper
• Identification and characterization of a mammalian protein interacting with 20S proteasome precursors.

  2000cited by this paper
• Identification of proteassemblin, a mammalian homologue of the yeast protein, Ump1p, that is required for normal proteasome assembly.

  2000cited by this paper
• Characterisation of the newly identified human Ump1 homologue POMP and analysis of LMP7(beta 5i) incorporation into 20 S proteasomes.

  2000cited by this paper
• Ump1p is required for proper maturation of the 20S proteasome and becomes its substrate upon completion of the assembly.

  1998cited by this paper
• Structure of 20S proteasome from yeast at 2.4Å resolution

  1997cited by this paper
• Autocatalytic subunit processing couples active site formation in the 20S proteasome to completion of assembly.

  1996cited by this paper
• Crystal structure of the 20S proteasome from the archaeon T. acidophilum at 3.4 A resolution.

  1995cited by this paper
• Quantitative analysis of protein far UV circular dichroism spectra by neural networks.

  1992cited by this paper
• Bioinformatics Original Paper Integrating Multi-attribute Similarity Networks for Robust Representation of the Protein Space

  year unknowncited by this paper

• Structural roles of Ump1 and β-subunit propeptides in proteasome biogenesis

  2024cites this paper
• Mechanochemical forces regulate the composition and fate of stalled nascent chains

  2024cites this paper
• Visualizing chaperone-mediated multistep assembly of the human 20S proteasome

  2024cites this paper
• Limiting 20S proteasome assembly leads to unbalanced nucleo-cytoplasmic distribution of 26S/30S proteasomes and chronic proteotoxicity

  2024cites this paper
• Chaperone-mediated assembly of the proteasome core particle - recent developments and structural insights.

  2022cites this paper
• Effective strategies on heterologous expression of plant heterotrimeric G-protein γ subunits without Gβ subunit partners.

  2022cites this paper
• Interaction with the Assembly Chaperone Ump1 Promotes Incorporation of the β7 Subunit into Half-Proteasome Precursor Complexes Driving Their Dimerization

  2022cites this paper
• A sequence‐based foldability score combined with AlphaFold2 predictions to disentangle the protein order/disorder continuum

  2022cites this paper
• Intracellular Dynamics of the Ubiquitin-Proteasome-System [ version 2 ; peer review : 3 approved ]

  2022cites this paper
• Intracellular localization of the proteasome in response to stress conditions

  2022cites this paper
• Structures of chaperone-associated assembly intermediates reveal coordinated mechanisms of proteasome biogenesis

  2021cites this paper
• Dynamic Regulation of the 26S Proteasome: From Synthesis to Degradation

  2019cites this paper
• Nuclear Transport of Yeast Proteasomes

  2019cites this paper
• Structural insights on the dynamics of proteasome formation

  2018cites this paper
• Gene Expression of Pneumocystis murina after Treatment with Anidulafungin Results in Strong Signals for Sexual Reproduction, Cell Wall Integrity, and Cell Cycle Arrest, Indicating a Requirement for Ascus Formation for Proliferation

  2018cites this paper
• Proteasome Structure and Assembly.

  2017cites this paper
• Putting it all together: intrinsic and extrinsic mechanisms governing proteasome biogenesis

  2017cites this paper
• Proteasome dynamics between proliferation and quiescence stages of Saccharomyces cerevisiae

  2016cites this paper
• Intracellular Dynamics of the Ubiquitin-Proteasome-System

  2015cites this paper
• Intracellular Dynamics of the Ubiquitin-Proteasome-System.

  2015cites this paper
• Functional assessment of intrinsic disorder central domains of BRCA1

  2015cites this paper
• Proteasome assembly from 15S precursors involves major conformational changes and recycling of the Pba1–Pba2 chaperone

  2015cites this paper
• The intrinsically disordered Sem1 protein functions as a molecular tether during proteasome lid biogenesis.

  2014cites this paper
• Snapshots of proteasomal precursor complexes reveal chaperone involvement in 20S proteasome biogenesis

  2014cites this paper
• Proteasome assembly

  2014cites this paper