Vertebrate tankyrase domain structure and sterile alpha motif (SAM)-mediated multimerization.
Tankyrases 1 and 2 are two highly related poly(ADP-ribose) polymerases that interact with a variety of cytoplasmic and nuclear proteins. Both proteins have been implicated in telomere length regulation, insulin signalling and centrosome function. To learn more about their mode of action, we have isolated the chicken tankyrase homologues and examined their interaction partners and subcellular location. Cross-species sequence comparison indicated that tankyrase domain structure is highly conserved and supports division of the ankyrin domain into five subdomains, which are each separated by a highly conserved LLEAAR/K motif. Glutathione S-transferase pull-down experiments demonstrated that the ankyrin domains of both proteins interact with chicken telomere repeat factor 1 (TRF1). Analysis of total cellular and nuclear proteins revealed that cells contain approximately twice as much tankyrase 1 as tankyrase 2. Although > or = 90% of each protein is present in the cytoplasm, both tankyrase 1 and 2 were detected in the nucleus. The nuclear location together with its ability to interact with TRF1, point to tankyrase 2 having a telomeric function. Yeast two-hybrid and cross-linking experiments show that both tankyrases can multimerize through their sterile-alpha motif domains. These results indicate that tankyrases may be master scaffolding proteins, capable of regulating assembly of large protein complexes.
De Rycker
Manu
M
Deparment of Molecular Genetics, Biochemistry and Microbiology, College of Medicine, University of Cincinnati, ML0524, 231 Albert Sabin Way, Cincinnati, OH 45267, U.S.A.
Venkatesan
Ranga N
RN
Wei
Chao
C
Price
Carolyn M
CM
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D017124:Conserved Sequence; D019281:Dimerization; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D038501:Tankyrases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D035321:Telomeric Repeat Binding Protein 1 / Q000378:metabolism
D038501:Tankyrases; D035321:Telomeric Repeat Binding Protein 1; C437334:TNKS2 protein, human; C418109:TNKS protein, human