J Am Soc Nephrol. 2004 Jan;15(1):41-51.

The death domain of kidney ankyrin interacts with Fas and promotes Fas-mediated cell death in renal epithelia.External

Del Rio, M., Imam, A., DeLeon, M., Gomez, G., Mishra, J., Ma, Q., Parikh, S., Devarajan, P.,
--- - Department of Nephrology, Children's Hospital at Montefiore, Albert Einstein College of Medicine, Bronx, New York, USA.
Ankyrins are a ubiquitously expressed family of conserved proteins that mediate the linkage of integral membrane proteins such as transporters and channels with the underlying cytoskeleton. Ankyrins possess a conserved death domain, the functional significance of which has remained puzzling. In this study, the death domain of AnkG190, the isoform of ankyrin expressed in kidney tubules, was used as bait in a yeast two-hybrid screen to identify interacting partners. One of these interactions was with the proapoptotic molecule Fas. This was confirmed by coimmunoprecipitation, colocalization, and glutathione S-transferase pull-down assays in cultured renal epithelial (MDCK) cells. Site-directed mutagenesis of a conserved arginine (R1496 in AnkG190), previously shown to be critical for the binding of Fas (R234 in Fas) to FADD, abolished the interaction of ankyrin's death domain with Fas. Overexpression of constructs containing ankyrin's death domain promoted Fas-mediated apoptosis in MDCK cells. The linkage between ankyrin and Fas was confirmed in vivo in mouse kidney tubule cells by coimmunoprecipitation and colocalization. In an established mouse model of renal ischemia-reperfusion injury characterized by apoptotic tubule cell death, the expression of both ankyrin and Fas was markedly induced, and the interaction between these molecules remained intact. The results identify a novel tethering interaction between ankyrin and Fas in kidney epithelia and suggest that AnkG190 may play a role as an adapter molecule in renal tubule cell death.
PMID: 14694156External
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vitro interaction
  DD1 DD2 Reference
Family DD1 DD2 Method Species Region Expression Species Region Expression
DD ANK3 Link FADD GST fusion protein pull-down Rat Not specified Not specified Not specified Not specified Not specified 14694156
DD ANK3 Link Fas GST fusion protein pull-down Rat Not specified Not specified Not specified Not specified Not specified 14694156
DD ANK3 Link Fas Yeast two-hybrid Rat Not specified Yeast Not specified Not specified Yeast 14694156
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vivo interaction
  Endogenous
expression
Overexpression DD1 DD2 Reference
Family DD1 DD2 Method Species Region Species Region
DD ANK3 Link FADD Co-Immunofluorescence staining MDCK Rat Not specified Not specified Not specified 14694156
DD ANK3 Link FADD Co-immunoprecipitation MDCK Rat Not specified Not specified Not specified 14694156
DD ANK3 Link Fas Co-Immunofluorescence staining MDCK Rat Not specified Not specified Not specified 14694156
DD ANK3 Link Fas Co-immunoprecipitation MDCK Rat Not specified Not specified Not specified 14694156
(Link: click this icon to show interactions only between the two corresponding DDs)