Development of a toolbox to dissect host-endosymbiont interactions and protein trafficking in the trypanosomatid Angomonas deanei.
Bacterial endosymbionts are found across the eukaryotic kingdom and profoundly impacted eukaryote evolution. In many endosymbiotic associations with vertically inherited symbionts, highly complementary metabolic functions encoded by host and endosymbiont genomes indicate integration of metabolic processes between the partner organisms. While endosymbionts were initially expected to exchange only metabolites with their hosts, recent evidence has demonstrated that also host-encoded proteins can be targeted to the bacterial symbionts in various endosymbiotic systems. These proteins seem to participate in regulating symbiont growth and physiology. However, mechanisms required for protein targeting and the specific endosymbiont targets of these trafficked proteins are currently unexplored owing to a lack of molecular tools that enable functional studies of endosymbiotic systems.



Here we show that the trypanosomatid Angomonas deanei, which harbors a β-proteobacterial endosymbiont, is readily amenable to genetic manipulation. Its rapid growth, availability of full genome and transcriptome sequences, ease of transfection, and high frequency of homologous recombination have allowed us to stably integrate transgenes into the A. deanei nuclear genome, efficiently generate null mutants, and elucidate protein localization by heterologous expression of a fluorescent protein fused to various putative targeting signals. Combining these novel tools with proteomic analysis was key for demonstrating the routing of a host-encoded protein to the endosymbiont, suggesting the existence of a specific endosymbiont-sorting machinery in A. deanei.



After previous reports from plants, insects, and a cercozoan amoeba we found here that also in A. deanei, i.e. a member of a fourth eukaryotic supergroup, host-encoded proteins can be routed to the bacterial endosymbiont. This finding adds further evidence to our view that the targeting of host proteins is a general strategy of eukaryotes to gain control over and interact with a bacterial endosymbiont. The molecular resources reported here establish A. deanei as a time and cost efficient reference system that allows for a rigorous dissection of host-symbiont interactions that have been, and are still being shaped over evolutionary time. We expect this system to greatly enhance our understanding of the biology of endosymbiosis.
Department of Biology, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany.
Jorge
J
Morales
Department of Biology, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany.
Sofia
S
Kokkori
Department of Biology, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany.
Diana
D
Weidauer
Plant Genome Group, DOE Joint Genome Institute, 2800 Mitchell Drive, 94598, Walnut Creek, CA, USA.
Jarrod
J
Chapman
Plant Genome Group, DOE Joint Genome Institute, 2800 Mitchell Drive, 94598, Walnut Creek, CA, USA.
Eugene
E
Goltsman
Plant Genome Group, DOE Joint Genome Institute, 2800 Mitchell Drive, 94598, Walnut Creek, CA, USA.
Daniel
D
Rokhsar
Department of Plant Biology, Carnegie Institution for Science, 260 Panama Street, 94305, Stanford, CA, USA.
Arthur R
AR
Grossman
Department of Biology, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany. e.nowack@uni-duesseldorf.de.
Eva C M
EC
Nowack
D000818:Animals; D001483:Base Sequence; D020562:Betaproteobacteria / Q000187:drug effects / Q000378:metabolism; D002934:Cinnamates / Q000494:pharmacology; D005822:Genetic Vectors / Q000378:metabolism; D018503:Genome, Protozoan; D023281:Genomics / Q000379:methods*; D005839:Gentamicins / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D059765:Homologous Recombination / Q000187:drug effects / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D016254:Mutagenesis, Insertional / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects; D015800:Protozoan Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013559:Symbiosis / Q000187:drug effects / Q000235:genetics; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics; D014351:Trypanosomatina / Q000187:drug effects / Q000235:genetics* / Q000382:microbiology*
D002934:Cinnamates; D005839:Gentamicins; D015800:Protozoan Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A; C010680:antibiotic G 418
Bacterial endosymbiont; Endosymbiosis; Homologous recombination; Protein targeting; Protist; Trypanosomatid