Dynamics of repeat-associated plasticity in the aaap gene family in Anaplasma marginale.

Anaplasmosis, the most prevalent tick-transmitted disease of cattle, is caused by the rickettsial intracellular parasite Anaplasma marginale. The pathogen replicates within a parasitophorous vacuole formed from the invagination of the erythrocyte membrane. Several strains of A. marginale form "tails" or "appendages" which are attached to, and extend out from, the cytoplasmic side of the parasitophorous vacuole. Genomic analysis of the parasite antigen distributed along the appendage led to the discovery of the aaap (Anaplasma appendage associated protein) gene family located within a highly plastic region in the genome. The aaap gene family consists of aaap and several alps (for aaap-like proteins), depending on the strain. These genes/proteins are characterized by repeat sequences. To investigate locus plasticity, different versions of the locus were cloned from the same strain as well as from different strains, sequenced and aligned to identify changes. Our findings show that repeat sequences both within and between genes facilitated rearrangement events within the locus. Structural variation of the locus in the St. Maries strain was further investigated during infection of different cellular environments, i.e., bovine erythrocytes and tick cells, with a reduction in subpopulations of the aaap locus within the tick as compared to erythrocytes. Interestingly, subpopulations bearing alternative locus structures began to arise again when the pathogen was transferred from the tick environment into a naïve calf. Additionally, the Aaap protein expression profile between blood and tick samples showed a regulatory shift, indicating a host-specific response. Alignment of the protein sequences from different species of Anaplasma reveals six similar repeating motifs that appear to be unique to a few species of Anaplasma. The role the aaap locus may play in the pathogenesis of the bovine host or in tick infection/transmission remains unknown; however, the changes in aaap locus subpopulations, locus structure, and protein expression indicate that these genes have a role in strain diversification.