Anaplasma phagocytophilum AnkA is tyrosine-phosphorylated at EPIYA motifs and recruits SHP-1 during early infection

IJdo, Jacob W.; Carlson, Adam C.; Kennedy, Elizabeth Lapovsky

doi:10.1111/j.1462-5822.2006.00871.x

Cited by 91 publications

(102 citation statements)

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“…AnkA sequences of all four clusters contained at their C-terminal end several tyrosine kinase phosphorylation sites predicted by Scansite (http: //scansite.mit.edu/). Some of these motifs have been shown also experimentally to be phosphorylated by the host cell kinase Src (31). However, only sequences belonging to clusters I and IV showed an extensive multiplication of the tyrosine phosphorylation sites due to the duplication of degenerated direct repeats (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Although the exact function of AnkA has not been determined yet (50), its C-terminal end has been shown to be secreted by the VirB/ VirD4-dependent type IV secretion system (T4SS) of Agrobacterium tumefaciens (34) and the Dot/Icm type T4SS of Legionella pneumophila (29). After translocation, AnkA is tyrosine phosphorylated by the host cell tyrosine kinases Src (31) and Abl (34). After phosphorylation, AnkA binds via SH2 domains to the host cell phosphatase SHP-1 (31), thus probably disturbing host cell signaling (52).…”

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confidence: 99%

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Distinct Host Species Correlate with Anaplasma phagocytophilum ankA Gene Clusters

et al. 2011

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Anaplasma phagocytophilum is a Gram-negative, tick-transmitted, obligate intracellular bacterium that elicits acute febrile diseases in humans and domestic animals. In contrast to the United States, human granulocytic anaplasmosis seems to be a rare disease in Europe despite the initial recognition of A. phagocytophilum as the causative agent of tick-borne fever in European sheep and cattle. Considerable strain variation has been suggested to occur within this species, because isolates from humans and animals differed in their pathogenicity for heterologous hosts. In order to explain host preference and epidemiological diversity, molecular characterization of A. phagocytophilum strains has been undertaken. Most often the 16S rRNA gene was used, but it might be not informative enough to delineate distinct genotypes of A. phagocytophilum. Previously, we have shown that A. phagocytophilum strains infecting Ixodes ricinus ticks are highly diverse in their ankA genes. Therefore, we sequenced the 16S rRNA and ankA genes of 194 A. phagocytophilum strains from humans and several animal species. Whereas the phylogenetic analysis using 16S rRNA gene sequences was not meaningful, we showed that distinct host species correlate with A. phagocytophilum ankA gene clusters.

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Section: Resultsmentioning

confidence: 99%

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Distinct Host Species Correlate with Anaplasma phagocytophilum ankA Gene Clusters

et al. 2011

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“…However, Wolbachia are unevenly distributed in adult tissues (Dobson, 2003;Ijdo et al, 2007;McGraw et al, 2002).…”

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confidence: 99%

Symmetric and asymmetric mitotic segregation patterns influence Wolbachia distribution in host somatic tissue

Albertson

Casper-Lindley

Cao

et al. 2009

Journal of Cell Science

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Wolbachia are maternally inherited bacterial endosymbionts that occupy many but not all tissues of adult insects. During the initial mitotic divisions in Drosophila embryogenesis, Wolbachia exhibit a symmetric pattern of segregation. Wolbachia undergo microtubule-dependent and cell-cycle-regulated movement between centrosomes. Symmetric segregation occurs during late anaphase when Wolbachia cluster around duplicated and separating centrosomes. This centrosome association is microtubule-dependent and promotes an even Wolbachia distribution throughout the host embryo. By contrast, during the later embryonic and larval neuroblast divisions, Wolbachia segregate asymmetrically with the apical self-renewing neuroblast. During these polarized asymmetric neuroblast divisions, Wolbachia colocalize with the apical centrosome and apically localized Par complex. This localization depends on microtubules, but not the cortical actin-based cytoskeleton. We also found that Wolbachia concentrate in specific regions of the adult brain, which might be a direct consequence of the asymmetric Wolbachia segregation in the earlier neuroblast divisions. Finally, we demonstrate that the fidelity of asymmetric segregation to the self-renewing neuroblast is lower in the virulent Popcorn strain of Wolbachia.

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“…Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacterial pathogens (14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26). These effectors contain one or more ankyrin repeats, each of which consists of a 33-amino-acid motif that mediates proteinprotein interactions (15).…”

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Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs

et al. 2015

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A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions. Orientia tsutsugamushi, an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here, in silico and manual sequence analyses revealed that a large proportion of O. tsutsugamushi strain Ikeda Anks bear a eukaryotic/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and/or endogenous SCF1 components. When coexpressed with FLAGAnk4_01 or FLAG-Ank9, a glutathione S-transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated O. tsutsugamushi-derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning O. tsutsugamushi, poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that O. tsutsugamushi strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an O. tsutsugamushi Ank does so during infection, and advance overall understanding of microbial F-box proteins. IMPORTANCEAnkyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets. Orientia tsutsugamushi, which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that O. tsutsugamushi strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an Orientia-derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide Orientia effector and the first example of an endogenous F-box-containing Ank-mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function.T he obligate intracellular bacterium Orientia tsutsugamushi is the causative agent of scrub typhus, a potentially fatal disease transmitted to humans through the bite of an infected trombiculid mite. O. tsutsugamushi invades phagocytic cells of the immune system, as well as endothelial cells of the skin and major organs (1-4). Di...

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Anaplasma phagocytophilum AnkA is tyrosine-phosphorylated at EPIYA motifs and recruits SHP-1 during early infection

Abstract: SummaryAnaplasma phagocytophilum is an intracellular pathogen that infects and survives in neutrophilic granulocytes. The A. phagocytophilum genome encodes a type four secretion system (T4SS) that

Cited by 91 publications

References 55 publications

Distinct Host Species Correlate with Anaplasma phagocytophilum ankA Gene Clusters

Distinct Host Species Correlate with Anaplasma phagocytophilum ankA Gene Clusters

Symmetric and asymmetric mitotic segregation patterns influence Wolbachia distribution in host somatic tissue

Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs

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