Anaplasma phagocytophilumModulates gp91phoxGene Expression through Altered Interferon Regulatory Factor 1 and PU.1 Levels and Binding of CCAAT Displacement Protein

Abstract

Infection of neutrophil precursors withAnaplasma phagocytophilum, the causative agent of human granulocytic ehrlichiosis, results in downregulation of the gp91^phoxgene, a key component of NADPH oxidase. We now show that repression of gp91^phoxgene transcription is associated with reduced expression of interferon regulatory factor 1 (IRF-1) and PU.1 in nuclear extracts ofA. phagocytophilum-infected cells. Loss of PU.1 and IRF-1 correlated with increased binding of the repressor, CCAAT displacement protein (CDP), to the promoter of the gp91^phoxgene. Reduced protein expression of IRF-1 was observed with or without gamma interferon (IFN-γ) stimulation, and the defect in IFN-γ signaling was associated with diminished binding of phosphorylated Stat1 to the Stat1 binding element of the IRF-1 promoter. The diminished levels of activator proteins and enhanced binding of CDP account for the transcriptional inhibition of the gp91^phoxgene duringA. phagocytophiluminfection, providing evidence of the first molecular mechanism that a pathogen uses to alter the regulation of genes that contribute to an effective respiratory burst.