| Abstract: |
Tankyrase is an ankyrin repeat-containing poly(ADP-ribose) polymerase originally isolated as a binding partner for the telomeric protein TRF1, but recently identified as a mitogen-activated protein kinase substrate implicated in regulation of Golgi vesicle trafficking. In this study, a novel human tankyrase, designated tankyrase 2, was isolated in a yeast two-hybrid screen as a binding partner for the Src homology 2 domain-containing adaptor protein Grb14. Tankyrase 2 is a 130-kDa protein, which lacks the N-terminal histidine/proline/serine-rich region of tankyrase, but contains a corresponding ankyrin repeat region, sterile α motif module, and poly(ADP-ribose) polymerase homology domain. The TANKYRASE 2 gene localizes to chromosome 10q23. 2 and is widely expressed, with mRNA transcripts particularly abundant in skeletal muscle and placenta. Upon subcellular fractionation, both Grb14 and tankyrase 2 associate with the low density microsome fraction, and association of these proteins in vivo can be detected by co-immunoprecipitation analysis. Deletion analyses implicate the N-terminal 110 amino acids of Grb14 and ankyrin repeats 10-19 of tankyrase 2 in mediating this interaction. This study supports a role for the tankyrases in cytoplasmic signal transduction pathways and suggests that vesicle trafficking may be involved in the subcellular localization or signaling function of Grb14. |
| Keywords: |
signal transduction; mitogen activated protein kinase; controlled study; unclassified drug; human cell; genetics; molecular genetics; protein motif; proteins; metabolism; gene; in situ hybridization, fluorescence; serine; cell protein; cell line; protein; in vivo study; protein interaction; enzyme substrate; gene library; molecular cloning; cloning, molecular; immunology; chemistry; fluorescence in situ hybridization; transcription regulation; amino acid sequence; molecular sequence data; sequence homology, amino acid; hybrid protein; amino terminal sequence; recombinant fusion proteins; genetic engineering; enzyme analysis; saccharomyces cerevisiae; sequence alignment; nucleotide sequence; glutathione transferase; immunoprecipitation; adaptor proteins, signal transducing; cellular distribution; binding site; cell fractionation; binding sites; sequence homology; fractionation; two hybrid system; biochemistry; isolation and purification; nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase; signal transducing adaptor protein; histidine; placenta; telomeric repeat binding factor 1; gene location; skeletal muscle; deletion mutant; chromosome map; chromosome mapping; proline; chromosome 10q; amino acid analysis; golgi complex; membrane vesicle; rna analysis; poly(adp-ribose) polymerases; adaptor protein; chromosome 10; affinity chromatography; chromatography, affinity; tankyrase; poly(adenosine diphosphate ribose); ankyrin; enzyme isolation; microsome; chromosomes, human, pair 10; humans; human; priority journal; article; tankyrases; src homology 2 domain; src homology domain; tnks protein, human; adaptor protein grb14; tankyrase 2; telomere protein; grb14 protein, human; tankyrase 2 gene
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