Structural analysis of a p44lmsp2 expression site of Anaplasma phagocytophilum in naturally infected ticks in Japan

Anaplasma phagocytophilum, an agent of human granulocytic anaplasmosis, infects neutrophils and causes an emerging tickborne febrile disease. The genome of this bacterium contains a large number of p44lmsp2-related genes encoding 44 kDa major outer-membrane proteins, and it is known that a specific p44lmsp2 gene is predominantly transcribed from a single expression locus. This study successfully characterized the genomic expression site for p44lmsp2 (3.8 kb) in uncultured A. phagocytophilum from Ixodes persulcatus ticks inhabiting a northern part of Japan. Comparative analysis of the sequences revealed that the structures of the expression sites in Japanese A. phagocytophilum were similar to those of US strains from human patients and European strains from a dog and sheep, but omp-1N (upstream from p44lmsp2) and a truncated recA (downstream from p44lmsp2) in the p44lmsp2 expression site seemed to share similarities with those of US and European strains. The central hypervariable region sequences of Japanese p44lmsp2 were found to be quite diverse (24.4-100 % amino acid similarities) and distinct from their closest relatives from US human patients or animal host origins (56.3-97.6% amino acid similarities) with some exceptions. Thus, this study provides significant information about the molecular characteristics of A. phagocytophilum in East Asia, as well as the global diversity of p44lmsp2.