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PMID:9003776

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Citation

Roehl, H, Bosenberg, M, Blelloch, R and Kimble, J (1996) Roles of the RAM and ANK domains in signaling by the C. elegans GLP-1 receptor. EMBO J. 15:7002-12

Abstract

In Caenorhabditis elegans, the GLP-1 receptor acts with a downstream transcriptional regulator, LAG-1, to mediate intercellular signaling. GLP-1 and LAG-1 are homologs of Drosophila Notch and Su(H) respectively. Here, we investigate the functions of two regions of the GLP-1 intracellular domain: the ANK repeat domain, which includes six cdc10/ankyrin repeats plus flanking amino acids, and the RAM domain, which spans approximately 60 amino acids just inside the transmembrane domain. First, we demonstrate that both ANK and RAM domains interact with the LAG-1 transcription factor. The interaction between the ANK domain and LAG-1 is only observed in nematodes by a co-localization assay and, therefore, may be either direct or indirect. By contrast, the interaction between the RAM domain and LAG-1 is likely to be direct, since it is observed by co-precipitation of the proteins in vitro as well as by yeast two-hybrid experiments. Second, we demonstrate that the RAM domain, when expressed in nematodes without a functional ANK repeat domain, does not mimic the unregulated receptor in directing cell fates or interfere with signaling by endogenous components. Finally, we show in yeast that the ANK repeats are strong transcriptional activators. Furthermore, missense mutations that eliminate receptor activity also abolish transcriptional activation by the GLP-1 ANK repeats in yeast. We speculate that one possible function for the ANK repeat domain is to act as a transcriptional co-activator with LAG-1.

Links

PubMed PMC452526

Keywords

Amino Acid Sequence; Animals; Animals, Genetically Modified; Caenorhabditis elegans/metabolism; Caenorhabditis elegans Proteins; DNA-Binding Proteins/metabolism; Helminth Proteins/metabolism; Molecular Sequence Data; Protein Binding; Receptors, Glucagon/chemistry; Receptors, Glucagon/metabolism; Saccharomyces cerevisiae/genetics; Sequence Homology, Amino Acid; Signal Transduction; Transcriptional Activation

Significance

Annotations

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

CAEEL:GLP1

enables

GO:0003713: transcription coactivator activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:GLP1

involved_in

GO:0045944: positive regulation of transcription by RNA polymerase II

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:GLP1

enables

GO:0061629: RNA polymerase II-specific DNA-binding transcription factor binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:V6CLJ5

F

Seeded From UniProt

complete

CAEEL:LAG1

located_in

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CAEEL:LAG1

involved_in

GO:0007219: Notch signaling pathway

ECO:0000353: physical interaction evidence used in manual assertion

WB:WBGene00001609

P

Seeded From UniProt

complete

CAEEL:LAG1

enables

GO:0005112: Notch binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P13508

F

Seeded From UniProt

complete

CAEEL:LAG1

enables

GO:0003690: double-stranded DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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