Truncation of the TPR domain of OGT alters substrate and glycosite selection

The auxilin family of J-domain proteins load Hsp70 onto clathrin-coated vesicles (CCVs) to drive uncoating. In vitro, auxilin function requires its ability to bind clathrin and stimulate Hsp70 ATPase activity via its J-domain. To test these requirements in vivo, we performed a mutational analysis of Swa2p, the yeast auxilin ortholog. Swa2p is a modular protein with three N-terminal clathrin-binding (CB) motifs, a ubiquitin association (UBA) domain, a tetratricopeptide repeat (TPR) domain, and a C-terminal J-domain. In vitro, clathrin binding is mediated by multiple weak interactions, but a Swa2p truncation lacking two CB motifs and the UBA domain retains nearly full function in vivo. Deletion of all CB motifs strongly abrogates clathrin disassembly but does not eliminate Swa2p function in vivo. Surprisingly, mutation of the invariant HPD motif within the J-domain to AAA only partially affects Swa2p function. Similarly, a TPR point mutation (G388R) causes a modest phenotype. However, Swa2p function is abolished when these TPR and J mutations are combined. The TPR and J-domains are not functionally redundant because deletion of either domain renders Swa2p nonfunctional. These data suggest that the TPR and J-domains collaborate in a bipartite interaction with Hsp70 to regulate its activity in clathrin disassembly.

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The Bub1-TPR Domain Interacts Directly with Mad3 to Generate Robust Spindle Checkpoint Arrest

Current Biology ◽

10.1016/j.cub.2019.06.011 ◽

2019 ◽

Vol 29 (14) ◽

pp. 2407-2414.e7 ◽

Cited By ~ 1

Author(s):

Ioanna Leontiou ◽

Nitobe London ◽

Karen M. May ◽

Yingrui Ma ◽

Lucile Grzesiak ◽

...

Keyword(s):

Spindle Checkpoint ◽

Tpr Domain

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Escherichia coli BepA has proteolytic and chaperone-like functions and acts in the degradation and biogenesis of β-barrel outer membrane proteins (OMP). The tetratricopeptide repeat (TPR) domain of BepA (orange) interacts with proteins of the β-barrel ass

Molecular Microbiology ◽

10.1111/mmi.13522 ◽

2017 ◽

Vol 106 (5) ◽

pp. i-i

Keyword(s):

Escherichia Coli ◽

Membrane Proteins ◽

Outer Membrane ◽

Outer Membrane Proteins ◽

Tetratricopeptide Repeat ◽

Tpr Domain

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Tcc1p, a Novel Protein Containing the Tetratricopeptide Repeat Motif, Interacts with Tup1p To Regulate Morphological Transition and Virulence in Candida albicans

Eukaryotic Cell ◽

10.1128/ec.00151-06 ◽

2006 ◽

Vol 5 (11) ◽

pp. 1894-1905 ◽

Cited By ~ 14

Author(s):

Aki Kaneko ◽

Takashi Umeyama ◽

Yuki Utena-Abe ◽

Satoshi Yamagoe ◽

Masakazu Niimi ◽

...

Keyword(s):

Candida Albicans ◽

Fungal Pathogen ◽

Affinity Purification ◽

Specific Protein ◽

Tetratricopeptide Repeat ◽

Human Fungal Pathogen ◽

The Common ◽

Transcriptional Gene Regulation ◽

Tpr Domain ◽

Novel Protein

ABSTRACT The transcriptional factor CaTup1p represses many genes involved in intracellular processes, including the yeast-hypha transition, in the human fungal pathogen Candida albicans. Using tandem affinity purification technology, we identified a novel protein that interacts with CaTup1p, named Tcc1p (Tup1p complex component). Tcc1p is a C. albicans-specific protein with a 736-amino-acid polypeptide with four tetratricopeptide repeat (TPR) motifs in the N-terminal portion. Tcc1p formed a protein complex with CaTup1p via the TPR domain of Tcc1p, independently of CaSsn6p-CaTup1p The tcc1Δ disruptant showed filamentous growth under conditions inducing the yeast form, as is true of the Catup1Δ mutant. Consistent with this result, the common set of hypha-specific genes was negatively regulated by both TCC1 and CaTUP1. These observations will provide new insights into CaTup1p-dependent transcriptional gene regulation in C. albicans.

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Human U4/U6 snRNP Recycling Factor p110: Mutational Analysis Reveals the Function of the Tetratricopeptide Repeat Domain in Recycling

Molecular and Cellular Biology ◽

10.1128/mcb.24.17.7392-7401.2004 ◽

2004 ◽

Vol 24 (17) ◽

pp. 7392-7401 ◽

Cited By ~ 26

Author(s):

Jan Medenbach ◽

Silke Schreiner ◽

Sunbin Liu ◽

Reinhard Lührmann ◽

Albrecht Bindereif

Keyword(s):

Amino Acids ◽

Protein Interactions ◽

Rna Binding ◽

Mutational Analysis ◽

Protein A ◽

Tetratricopeptide Repeat ◽

Terminal Sequence ◽

Recognition Motifs ◽

Tpr Domain

ABSTRACT After each spliceosome cycle, the U4 and U6 snRNAs are released separately and are recycled to the functional U4/U6 snRNP, requiring in the mammalian system the U6-specific RNA binding protein p110 (SART3). Its domain structure is made up of an extensive N-terminal domain with at least seven tetratricopeptide repeat (TPR) motifs, followed by two RNA recognition motifs (RRMs) and a highly conserved C-terminal sequence of 10 amino acids. Here we demonstrate under in vitro recycling conditions that U6-p110 is an essential splicing factor. Recycling activity requires both the RRMs and the TPR domain but not the highly conserved C-terminal sequence. For U6-specific RNA binding, the two RRMs with some flanking regions are sufficient. Yeast two-hybrid assays reveal that p110 interacts through its TPR domain with the U4/U6-specific 90K protein, indicating a specific role of the TPR domain in spliceosome recycling. On the 90K protein, a short internal region (amino acids 416 to 550) suffices for the interaction with p110. Together, these data suggest a model whereby p110 brings together U4 and U6 snRNAs through both RNA-protein and protein-protein interactions.

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Distribution of Activator of G-Protein Signaling 3 within the Aggresomal Pathway: Role of Specific Residues in the Tetratricopeptide Repeat Domain and Differential Regulation by the AGS3 Binding Partners Giα and Mammalian Inscuteable

Molecular and Cellular Biology ◽

10.1128/mcb.01018-09 ◽

2010 ◽

Vol 30 (6) ◽

pp. 1528-1540 ◽

Cited By ~ 18

Author(s):

Ali Vural ◽

Sadik Oner ◽

Ningfei An ◽

Violaine Simon ◽

Dzwokai Ma ◽

...

Keyword(s):

G Protein ◽

G Protein Signaling ◽

Protein Signaling ◽

Single Nucleotide ◽

Signaling Systems ◽

Binding Partners ◽

Repeat Domain ◽

Tetratricopeptide Repeat Domain ◽

Tpr Domain

ABSTRACT AGS3, a receptor-independent activator of G-protein signaling, is involved in unexpected functional diversity for G-protein signaling systems. AGS3 has seven tetratricopeptide (TPR) motifs upstream of four G-protein regulatory (GPR) motifs that serve as docking sites for Giα-GDP. The positioning of AGS3 within the cell and the intramolecular dynamics between different domains of the proteins are likely key determinants of their ability to influence G-protein signaling. We report that AGS3 enters into the aggresome pathway and that distribution of the protein is regulated by the AGS3 binding partners Giα and mammalian Inscuteable (mInsc). Giα rescues AGS3 from the aggresome, whereas mInsc augments the aggresome-like distribution of AGS3. The distribution of AGS3 to the aggresome is dependent upon the TPR domain, and it is accelerated by disruption of the TPR organizational structure or introduction of a nonsynonymous single-nucleotide polymorphism. These data present AGS3, G-proteins, and mInsc as candidate proteins involved in regulating cellular stress associated with protein-processing pathologies.

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Molecular Recognition via Coupled Folding and Binding in a TPR Domain

Journal of Molecular Biology ◽

10.1016/j.jmb.2004.12.017 ◽

2005 ◽

Vol 346 (3) ◽

pp. 717-732 ◽

Cited By ~ 63

Author(s):

Matthew J. Cliff ◽

Mark A. Williams ◽

John Brooke-Smith ◽

David Barford ◽

John E. Ladbury

Keyword(s):

Molecular Recognition ◽

Tpr Domain

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Routing of Hansenula polymorpha Alcohol Oxidase: An Alternative Peroxisomal Protein-sorting Machinery

Molecular Biology of the Cell ◽

10.1091/mbc.e03-04-0258 ◽

2004 ◽

Vol 15 (3) ◽

pp. 1347-1355 ◽

Cited By ~ 53

Author(s):

Katja Gunkel ◽

Ralf van Dijk ◽

Marten Veenhuis ◽

Ida J. van der Klei

Keyword(s):

Amino Acids ◽

Protein Translocation ◽

Hansenula Polymorpha ◽

Alcohol Oxidase ◽

N Terminus ◽

Dependent Protein ◽

Peptides Synthesis ◽

Translocation Pathway ◽

Tpr Domain ◽

Fad Binding

Import of Hansenula polymorpha alcohol oxidase (AO) into peroxisomes is dependent on the PTS1 receptor, HpPex5p. The PTS1 of AO (-LARF) is sufficient to direct reporter proteins to peroxisomes. To study AO sorting in more detail, strains producing mutant AO proteins were constructed. AO containing a mutation in the FAD binding fold was mislocalized to the cytosol. This indicates that the PTS1 of AO is not sufficient for import of AO. AO protein in which the PTS1 was destroyed (-LARA) was normally sorted to peroxisomes. Moreover, C-terminal deletions of up to 16 amino acids did not significantly affect AO import, indicating that the PTS1 was not necessary for targeting. Consistent with these observations we found that AO import occurred independent from the C-terminal TPR-domain of HpPex5p, known to bind PTS1 peptides. Synthesis of the N-terminal domain (amino acids 1-272) of HpPex5p in pex5 cells restored AO import, whereas other PTS1 proteins were mislocalized to the cytosol. These data indicate that AO is imported via a novel HpPex5p-dependent protein translocation pathway, which does not require the PTS1 of AO and the C-terminal TPR domains of HpPex5p, but involves FAD binding and the N-terminus of HpPex5p.

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