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2011 ; 50
(51
): 11084-96
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Nondegradative ubiquitination of apoptosis inducing factor (AIF) by X-linked
inhibitor of apoptosis at a residue critical for AIF-mediated chromatin
degradation
#MMPMID22103349
Lewis EM
; Wilkinson AS
; Davis NY
; Horita DA
; Wilkinson JC
Biochemistry
2011[Dec]; 50
(51
): 11084-96
PMID22103349
show ga
Apoptosis inducing factor (AIF) is a mediator of caspase-independent cell death
that is also necessary for mitochondrial energy production. How these seemingly
opposite cellular functions of AIF are controlled is poorly understood. X-linked
inhibitor of apoptosis (XIAP) is an endogenous inhibitor of caspases that also
regulates several caspase-independent signaling pathways. The RING domain of XIAP
possesses E3 ubiquitin ligase activity, though the importance of this function to
signal regulation remains incompletely defined. XIAP binds and ubiquitinates AIF,
and in this study, we determined the functional consequences of XIAP-mediated AIF
ubiquitination. Unlike canonical ubiquitination, XIAP-dependent AIF
ubiquitination did not lead to proteasomal degradation of AIF. Experiments using
ubiquitin mutants demonstrated that the XIAP-dependent ubiquitin linkage was not
formed through the commonly used lysine 48, suggesting a noncanonical ubiquitin
linkage is employed. Further studies demonstrated that only lysine 255 of AIF was
a target of XIAP-dependent ubiquitination. Using recombinant AIF, we determined
that mutating lysine 255 of AIF interferes with the ability of AIF not only to
bind DNA but also to degrade chromatin in vitro. These data indicate that XIAP
regulates the death-inducing activity of AIF through nondegradative
ubiquitination, further defining the role of XIAP in controlling AIF and
caspase-independent cell death pathways.