Mechanisms Underlying the Delayed Activation of the Cap1 Transcription Factor in Candida albicans following Combinatorial Oxidative and Cationic Stress Important for Phagocytic Potency
Authors: Kos Iaroslava , Patterson Miranda J. , Znaidi Sadri , Kaloriti Despoina , da Silva Dantas Alessandra , Herrero-de-Dios Carmen M. , d’Enfert Christophe , Brown Alistair J. P. , Quinn Janet 
Published: March 2016

Following phagocytosis, microbes are exposed to an array of antimicrobial weapons that include reactive oxygen species (ROS) and cationic fluxes. This is significant as combinations of oxidative and cationic stresses are much more potent than the corresponding single stresses, triggering the synergistic killing of the fungal pathogen
Candida albicans
by “stress pathway interference.” Previously we demonstrated that combinatorial oxidative plus cationic stress triggers a dramatic increase in intracellular ROS levels compared to oxidative stress alone. Here we show that activation of Cap1, the major regulator of antioxidant gene expression in
C. albicans
, is significantly delayed in response to combinatorial stress treatments and to high levels of H
2
O
2
. Cap1 is normally oxidized in response to H
2
O
2
; this masks the nuclear export sequence, resulting in the rapid nuclear accumulation of Cap1 and the induction of Cap1-dependent genes. Here we demonstrate that following exposure of cells to combinatorial stress or to high levels of H
2
O
2
, Cap1 becomes trapped in a partially oxidized form, Cap1
OX-1
. Notably, Cap1-dependent gene expression is not induced when Cap1 is in this partially oxidized form. However, while Cap1
OX-1
readily accumulates in the nucleus and binds to target genes following high-H
2
O
2
stress, the nuclear accumulation of Cap1
OX-1
following combinatorial H
2
O
2
and NaCl stress is delayed due to a cationic stress-enhanced interaction with the Crm1 nuclear export factor. These findings define novel mechanisms that delay activation of the Cap1 transcription factor, thus preventing the rapid activation of the stress responses vital for the survival of
C. albicans
within the host.

IMPORTANCE
Combinatorial stress-mediated synergistic killing represents a new unchartered area in the field of stress signaling. This phenomenon contrasts starkly with “stress cross-protection,” where exposure to one stress protects against subsequent exposure to a different stress. Previously we demonstrated that the pathogen
Candida albicans
is acutely sensitive to combinations of cationic and oxidative stresses, because the induction of H
2
O
2
-responsive genes is blocked in the presence of cationic stress. We reveal that this is due to novel mechanisms that delay activation of the Cap1 AP-1-like transcription factor, the major regulator of the H
2
O
2
-induced regulon. Cap1 becomes trapped in a partially oxidized form following simultaneous exposure to oxidative and cationic stresses. In addition, cationic stress promotes the interaction of Cap1 with the Crm1 nuclear export factor, thus inhibiting its nuclear accumulation. These mechanisms probably explain the potency of neutrophils, which employ multiple stresses to kill fungal pathogens.