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PMID:14871887

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Citation

Secombe, J and Parkhurst, SM (2004) Drosophila Topors is a RING finger-containing protein that functions as a ubiquitin-protein isopeptide ligase for the hairy basic helix-loop-helix repressor protein. J. Biol. Chem. 279:17126-33

Abstract

Transcriptional repression plays an essential role in many aspects of metazoan development. Drosophila hairy is a primary pair-rule gene encoding a basic helix-loop-helix class transcriptional repressor that is required for proper segmentation. Previous characterization of Hairy-binding proteins has implicated two different classes of histone deacetylase as mediators of Hairy repression. Here, we present the characterization of a novel Hairy-interacting protein (dTopors) that binds specifically to the basic region of Hairy, but does not affect the ability of Hairy to bind DNA. By reducing the gene dose of dtopors, we demonstrate that it acts genetically as an antagonist of Hairy-mediated transcriptional repression. Consistent with this genetic interaction, we show that that recombinant dTopors protein possesses ubiquitin-protein isopeptide ligase activity in vitro and that dTopors mediates Hairy polyubiquitination and can lead to Hairy degradation. This work provides the first evidence that regulated proteolysis of Hairy is required for correct segmentation.

Links

PubMed Online version:10.1074/jbc.M310097200

Keywords

Amino Acid Sequence; Animals; Basic Helix-Loop-Helix Transcription Factors; Carrier Proteins/chemistry; Carrier Proteins/physiology; Cell Line; Cloning, Molecular; DNA/chemistry; DNA, Complementary/metabolism; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/physiology; Drosophila/metabolism; Drosophila Proteins; Gene Deletion; Glutathione Transferase/metabolism; Histone Deacetylases/metabolism; Humans; Insect Proteins/metabolism; Models, Genetic; Molecular Sequence Data; Neoplasm Proteins; Nuclear Proteins/chemistry; Nuclear Proteins/physiology; Protein Binding; Recombinant Proteins/metabolism; Repressor Proteins/metabolism; Sequence Homology, Amino Acid; Transcription Factors/chemistry; Transcription Factors/physiology; Transcription, Genetic; Ubiquitin/metabolism; Ubiquitin-Protein Ligases/chemistry; Zinc Fingers

Significance

Annotations

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

DROME:HAIR

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9V8P9

F

Seeded From UniProt

complete

HUMAN:ARL8A

involved_in

GO:0007059: chromosome segregation

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:Q9NVJ2

P

Seeded From UniProt

complete

HUMAN:ARL8B

involved_in

GO:0007059: chromosome segregation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:TOPRS

involved_in

GO:0000209: protein polyubiquitination

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:TOPRS

involved_in

GO:0006511: ubiquitin-dependent protein catabolic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:TOPRS

enables

GO:0061630: ubiquitin protein ligase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DROME:TOPRS

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P14003

F

Seeded From UniProt

complete

DROME:TOPRS

involved_in

GO:0006355: regulation of transcription, DNA-templated

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:P14003

P

Seeded From UniProt

complete

DROME:ARL8

involved_in

GO:0000819: sister chromatid segregation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

See also

References

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