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Papers In Press, published online ahead of print February 19, 2002
J. Biol. Chem, 10.1074/jbc.M112266200
Submitted on December 21, 2001
Revised on February 8, 2002
Accepted on February 19, 2002
Molecular Pathogenesis and Pathology, Skirball Institute, NYU School of Medicine, New York, NY 10016
Corresponding Author: smithsu{at}saturn.med.nyu.edu
Tankyrase1, a human telomeric poly(ADP-ribose) polymerase (PARP), was originally identified through its interaction with TRF1, a negative regulator of telomere length. Tankyrase1 ADP-ribosylates TRF1 in vitro and its overexpression induces telomere elongation in human cancer cells. In addition to its telomeric localization, tankyrase1 resides at multiple subcellular sites, suggesting additional functions for this protein. Here we identify TAB182, a novel tankyrase1-binding protein of 182 kD. TAB182 displays a complex pattern of subcellular localization. TAB182 localizes to the nucleus in a heterochromatic staining pattern and to the cytoplasm, where it costains with the cortical actin network. TAB182 coimmunoprecipitates with tankyrase1 from human cells and serves as an acceptor of poly(ADP-ribosyl)ation by tankyrase1 in vitro. Like TRF1, TAB182 binds to the ankyrin domain (comprising 24 ankyrin repeats) of tankyrase1. Surprisingly, dissection of this domain reveals multiple, discrete and overlapping, binding sites for TRF1 and TAB182. Thus, we demonstrate five well conserved ANK repeat clusters (ARCs) in tankyrase1. While each of the five ARCs independently binds to TRF1, only three of the five bind toTAB182. These findings suggest that tankyrase1 may act as a scaffold for large molecular weight complexes made up of multiple binding proteins. We discuss potential roles for tankyrase1-mediated higher order complexes at telomeres and at other subcellular sites.
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