UFD-2 is an adaptor-assisted E3 ligase targeting unfolded proteins

Hellerschmied, Doris; Roessler, Max; Lehner, Anita; Gazda, Linn; Stejskal, Karel; Imre, Richard; Mechtler, Karl; Dammermann, Alexander; Clausen, Tim

doi:10.1038/s41467-018-02924-7

Cited by 22 publications

(30 citation statements)

References 63 publications

(92 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We first studied two mutants deficient in myosin binding, namely a deletion mutant lacking the UCS domain (residues 1–521, ∆UCS) 14,24 and a variant carrying a point mutation (N758Y) in the myosin-binding canyon 19 . Deletion of the UCS domain had a strong impact on UNC-45 chaperone function, as seen by the reduced levels of soluble MHC-B in the insect cell co-expression system (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Subsequent studies in C. elegans , Xenopus, Zebrafish, and Drosophila confirmed the role of UNC-45 as a myosin-specific chaperone, promoting the folding of the myosin ATPase domain as well as coordinating the assembly of thick filaments during muscle development 14–19 . In higher organisms, UNC-45 is particularly important for the maturation of myosin II in muscle cells 20–22 and, together with Hsp90 (heat shock protein 90) and UFD-2 (ubiquitin fusion degradation 2), maintaining the functionality of myosin filaments during stress situations 23,24 . Mechanistic insight into the chaperone function of UNC-45 comes from structural studies revealing its 3-domain architecture 19,25 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin

et al. 2019

Self Cite

View full text Add to dashboard Cite

Myosin is a motor protein that is essential for a variety of processes ranging from intracellular transport to muscle contraction. Folding and assembly of myosin relies on a specific chaperone, UNC-45. To address its substrate-targeting mechanism, we reconstitute the interplay between Caenorhabditis elegans UNC-45 and muscle myosin MHC-B in insect cells. In addition to providing a cellular chaperone assay, the established system enabled us to produce large amounts of functional muscle myosin, as evidenced by a biochemical and structural characterization, and to directly monitor substrate binding to UNC-45. Data from in vitro and cellular chaperone assays, together with crystal structures of binding-deficient UNC-45 mutants, highlight the importance of utilizing a flexible myosin-binding domain. This so-called UCS domain can adopt discrete conformations to efficiently bind and fold substrate. Moreover, our data uncover the molecular basis of temperature-sensitive UNC-45 mutations underlying one of the most prominent motility defects in C. elegans.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…44 To determine if IV-FPOP can do so within a multiorgan system, we looked at the peptide-level oxidation pattern of the myosin chaperone protein UNC-45. Although only two modified peptides were observed for UNC-45, it was chosen for further study, because a crystal structure is available for the C. elegans form of this protein alone (PDB ID 5MZU(45)) and in complex with a heat shock protein 90 (Hsp90) peptide (PDB ID 4I2Z 46 ). Only multiresidue modified peptides were observed (Figure 5b), rendering the contribution of the different residues to the overall extent of modification indistinguishable.…”

Section: Resultsmentioning

confidence: 99%

Illuminating Biological Interactions with in Vivo Protein Footprinting

Espino

Jones

2019

Anal. Chem.

View full text Add to dashboard Cite

Protein footprinting coupled with mass spectrometry is being increasingly used for the study of protein interactions and conformations. The hydroxyl radical footprinting method, fast photochemical oxidation of proteins (FPOP), utilizes hydroxyl radicals to oxidatively modify solvent accessible amino acids. Here, we describe the further development of FPOP for protein structural analysis in vivo (IV-FPOP) with Caenorhabditis elegans. C. elegans, part of the nematode family, are used as model systems for many human diseases. The ability to perform structural studies in these worms would provide insight into the role of structure in disease pathogenesis. Many parameters were optimized for labeling within the worms including the microfluidic flow system and hydrogen peroxide concentration. IV-FPOP was able to modify several hundred proteins in various organs within the worms. The method successfully probed solvent accessibility similarily to in vitro FPOP, demonstrating its potential for use as a structural technique in a multiorgan system. The coupling of the method with mass spectrometry allows for amino-acid-residue-level structural information, a higher resolution than currently available in vivo methods.

show abstract

“…However, the predicted structure of the U-box is very similar to that of the RING finger domain [30], which indicates that U-box proteins may also have the capability to function as E3 ligases independently. Indeed, UFD2 can function as a bona fide E3 ubiquitin ligase to promote ubiquitin conjugation of unfolded proteins [31]. Thus, UBE4B and its homologs can clearly function as E3 ligases, and their E4 function may represent a specialized type of E3 activity with mono-or oligoubiquitinated proteins as substrates.…”

Section: K63-linked Polyubiquitination and Stabilization And Inhibitimentioning

confidence: 99%

The E3/E4 ubiquitin conjugation factor UBE4B interacts with and ubiquitinates the HTLV-1 Tax oncoprotein to promote NF-κB activation

Harhaj

Mohanty²,

Han

et al. 2020

Preprint

View full text Add to dashboard Cite

Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL), and the neurological disease HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-1 Tax protein persistently activates the NF-κB pathway to enhance the proliferation and survival of HTLV-1 infected T cells. Lysine 63 (K63)-linked polyubiquitination of Tax provides an important regulatory mechanism that promotes Tax-mediated interaction with the IKK complex and activation of NF-κB; however, the host proteins regulating Tax ubiquitination are largely unknown. To identify novel Tax interacting proteins that may regulate its ubiquitination we conducted a yeast two-hybrid screen using Tax as bait. This screen yielded the E3/E4 ubiquitin conjugation factor UBE4B as a novel binding partner for Tax.Here, we confirmed the interaction between Tax and UBE4B in mammalian cells by coimmunoprecipitation assays and demonstrated colocalization by proximity ligation assay and confocal microscopy. Overexpression of UBE4B specifically enhanced Tax-induced NF-κB activation, whereas knockdown of UBE4B impaired Tax-induced NF-κB activation and the induction of NF-B target genes in T cells and ATLL cell lines. Furthermore, depletion of UBE4B with shRNA resulted in apoptotic cell death and diminished the proliferation of ATLL cell lines. Finally, overexpression of UBE4B enhanced Tax polyubiquitination, and knockdown or CRISPR/Cas9-mediated knockout of UBE4B attenuated both K48-and K63-linked polyubiquitination of Tax. Collectively, these results implicate UBE4B in HTLV-1 Tax polyubiquitination and downstream NF-B activation. conjugation factor UBE4B as a novel Tax binding protein that promotes both K48-and K63-linked polyubiquitination of Tax. Knockdown or knockout of UBE4B impairs Taxinduced NF-B activation and triggers apoptosis of HTLV-1 transformed cells. Therefore, UBE4B is an integral host factor that supports HTLV-1 Tax polyubiquitination, NF-B activation and cell survival.

show abstract

UFD-2 is an adaptor-assisted E3 ligase targeting unfolded proteins

Cited by 22 publications

References 63 publications

Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin

Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin

Illuminating Biological Interactions with in Vivo Protein Footprinting

The E3/E4 ubiquitin conjugation factor UBE4B interacts with and ubiquitinates the HTLV-1 Tax oncoprotein to promote NF-κB activation

Contact Info

Product

Resources

About