BAIT

RSP5

MDP1, MUT2, NPI1, SMM1, UBY1, NEDD4 family E3 ubiquitin-protein ligase, L000001054, L000001779, L000001220, YER125W
E3 ubiquitin ligase of NEDD4 family; regulates many cellular processes including MVB sorting, heat shock response, transcription, endocytosis, ribosome stability; mutant tolerates aneuploidy; autoubiquitinates; ubiquitinates Sec23p and Sna3p; deubiquitinated by Ubp2p; activity regulated by SUMO ligase Siz1p, in turn regulates Siz1p SUMO ligase activity; required for efficient Golgi-to-ER trafficking in COPI mutants; human homolog implicated in Liddle syndrome
GO Process (33)
GO Function (3)
GO Component (9)

Gene Ontology Biological Process

Saccharomyces cerevisiae (S288c)
PREY

DIA1

YMR316W
Protein of unknown function; involved in invasive and pseudohyphal growth; green fluorescent protein (GFP)-fusion protein localizes to the cytoplasm in a punctate pattern
GO Process (2)
GO Function (0)
GO Component (1)

Gene Ontology Cellular Component

Saccharomyces cerevisiae (S288c)

Biochemical Activity (Ubiquitination)

An interaction is inferred from the biochemical effect of one protein upon another, for example, GTP-GDP exchange activity or phosphorylation of a substrate by a kinase. The bait protein executes the activity on the substrate hit protein. A Modification value is recorded for interactions of this type with the possible values Phosphorylation, Ubiquitination, Sumoylation, Dephosphorylation, Methylation, Prenylation, Acetylation, Deubiquitination, Proteolytic Processing, Glucosylation, Nedd(Rub1)ylation, Deacetylation, No Modification, Demethylation.

Publication

Ubiquitination screen using protein microarrays for comprehensive identification of Rsp5 substrates in yeast.

Gupta R, Kus B, Fladd C, Wasmuth J, Tonikian R, Sidhu S, Krogan NJ, Parkinson J, Rotin D

Ubiquitin-protein ligases (E3s) are responsible for target recognition and regulate stability, localization or function of their substrates. However, the substrates of most E3 enzymes remain unknown. Here, we describe the development of a novel proteomic in vitro ubiquitination screen using a protein microarray platform that can be utilized for the discovery of substrates for E3 ligases on a global scale. ... [more]

Mol. Syst. Biol. Jun. 07, 2007; 3(0);116 [Pubmed: 17551511]

Throughput

  • High Throughput|Low Throughput

Additional Notes

  • High Throughput: A ubiquitination assay was applied to a protein microarray in order to identify substrates of the E3 enzyme Rsp5.
  • Low Throughput: The in vitro reaction contained E1, Ubc4 [E2], and ubiquitin.

Related interactions

InteractionExperimental Evidence CodeDatasetThroughputScoreCurated ByNotes
RSP5 DIA1
Affinity Capture-MS
Affinity Capture-MS

An interaction is inferred when a bait protein is affinity captured from cell extracts by either polyclonal antibody or epitope tag and the associated interaction partner is identified by mass spectrometric methods.

Low-BioGRID
1516752
DIA1 RSP5
Negative Genetic
Negative Genetic

Mutations/deletions in separate genes, each of which alone causes a minimal phenotype, but when combined in the same cell results in a more severe fitness defect or lethality under a given condition. This term is reserved for high or low throughput studies with scores.

High-0.2555BioGRID
2063606
RSP5 DIA1
Protein-peptide
Protein-peptide

An interaction is detected between a protein and a peptide derived from an interaction partner. This includes phage display experiments.

High-BioGRID
570785
RSP5 DIA1
Reconstituted Complex
Reconstituted Complex

An interaction is detected between purified proteins in vitro.

High-BioGRID
238372
DIA1 RSP5
Two-hybrid
Two-hybrid

Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.

High-BioGRID
-
DIA1 RSP5
Two-hybrid
Two-hybrid

Bait protein expressed as a DNA binding domain (DBD) fusion and prey expressed as a transcriptional activation domain (TAD) fusion and interaction measured by reporter gene activation.

High-BioGRID
-

Curated By

  • BioGRID