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PMID:11717310

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Citation

Murillas, R, Simms, KS, Hatakeyama, S, Weissman, AM and Kuehn, MR (2002) Identification of developmentally expressed proteins that functionally interact with Nedd4 ubiquitin ligase. J. Biol. Chem. 277:2897-907

Abstract

Nedd4 is a HECT domain-containing ubiquitin ligase that mediates ubiquitylation and proteasome degradation of target proteins. The molecular basis for the interaction of Nedd4 with substrates lies in its WW domains, which can bind proline-rich (PY) domains in target proteins. Nedd4 is a developmentally expressed protein and may have a fundamental role to play in embryonic processes. However, whether Nedd4 has such a function is currently unknown, in part because few developmentally regulated ubiquitylation substrates have been identified or characterized. We have carried out a yeast two-hybrid screen and identified four proteins expressed in the mid-gestation embryo that are able to interact with Nedd4. Characterization of their functional interaction with Nedd4 in vitro and in vivo demonstrated that three of the four are bona fide Nedd4 binding partners, and two have the capacity to be ubiquitylation substrates. One of these is the first identified nonviral substrate for Nedd4-mediated monoubiquitylation. Interestingly, neither of these two ubiquitylated proteins interacts with Nedd4 through PY-mediated mechanisms. For one of the three Nedd4 binding partners, there was no discernable evidence of ubiquitylation. However, this protein clearly associates with Nedd4 through its PY domains and can alter the location of Nedd4 in cells, suggesting a role other than as a ubiquitylation substrate.

Links

PubMed Online version:10.1074/jbc.M110047200

Keywords

Blotting, Western; Calcium-Binding Proteins/metabolism; Cell Line; Cell Nucleus/metabolism; Cysteine Endopeptidases; DNA, Complementary/metabolism; Endosomal Sorting Complexes Required for Transport; Expressed Sequence Tags; Glutathione Transferase/metabolism; Humans; Ligases/metabolism; Microscopy, Fluorescence; Multienzyme Complexes/antagonists & inhibitors; Mutagenesis, Site-Directed; Plasmids/metabolism; Precipitin Tests; Proteasome Endopeptidase Complex; Protein Binding; Protein Biosynthesis; Protein Structure, Tertiary; Recombinant Fusion Proteins/metabolism; Transfection; Two-Hybrid System Techniques; Ubiquitin/metabolism; Ubiquitin-Protein Ligases

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:A0A0J9YUP5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:A0A0J9YUP5

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:F8VQG7

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:F8VQG7

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:NEDD4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

MGI:MGI:2684414
PR:Q6A037
PR:Q8C7U1

F

Seeded From UniProt

complete

MOUSE:NEDD4

enables

GO:0061630: ubiquitin protein ligase activity

ECO:0000314: direct assay evidence used in manual assertion

F

  • has_input:(MGI:MGI:2136825)|has_input:(MGI:MGI:2684414)

Seeded From UniProt

complete

MOUSE:Q69ZK1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:Q69ZK1

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:N4BP1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:N4BP1

located_in

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:Q8BX85

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:Q8BX85

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:N4BP3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P46935

F

Seeded From UniProt

complete

MOUSE:N4BP3

located_in

GO:0031410: cytoplasmic vesicle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:NEDD4

enables

GO:0061630: ubiquitin protein ligase activity

ECO:0000314: direct assay evidence used in manual assertion

F

has_direct_input:(MGI:MGI:2136825)|has_direct_input:(MGI:MGI:2684414)

Seeded From UniProt

complete


See also

References

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