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Lukes, J.

Publications and source records attributed to Lukes, J..

At least 19 recordsLinked to original sources

A subcellular spatial proteome of the non-photosynthetic euglenid Euglena longa

Euglena longa is a heterotrophic model euglenid, of significant research interest as a unicellular eukaryote. To provide broad insight into the localisation of its proteome we used subcellular spatial proteomics (SSP), based on a label-free version of LOPIT-DC (Localisation of Organelle Proteins by Isotope Tagging after Differential Centrifugation), in which organelles are separated by differential centrifugation and each protein is assigned to a compartment from its abundance profile across the fractions. We detected 5,909 proteins, using protein abundance profiles across fractions to resolve 16 different subcellular compartments, such as the plastid, pellicle, and peroxisomes, and assigned high-confidence localisations to 2,955 of them. This study serves as a proteomic dataset to resolve protein compartmentalisation for this euglenid, enabling evolutionary, biological and functional insight and analysis. We are drafting manuscripts concerning the metabolic nature of the plastid, the evolutionary origins of the pellicle, and the functions of the peroxisome. Investigators interested in using this data prior to publication are encouraged to contact us to avoid overlap and to facilitate coordinated and collaborative use of this resource.

cell biology↗

A threonine-to-aspartate ACA codon reassignment in uncultivated Acidimicrobiales bacteria

The list of alternative genetic codes continues to grow, yet sense-to-sense codon reassignments remain very rare. Here we analyze a new sense-to-sense codon reassignment in bacteria, where the ACA codon, which canonically specifies threonine, has been predicted to encode aspartate. This so far unprecedented genetic code alteration is confined to a clade within the RAAP-2 family of the order Acidimicrobiales (phylum Actinomycetota). Validated by multiple alignments of highly conserved proteins, the reassignment is further supported by the presence of a tRNAUGU that lacks the G1:C72 threonylation identity element, suggesting that this tRNA is charged with aspartate instead of threonine. Our findings not only expand the set of known natural departures from the canonical genetic code by a novel type of sense-to-sense reassignment but also implicate the ACA codon that was so far considered invariably conserved in its meaning.

genomics↗

Absence of endosymbiotic gene transfer in symbiont-bearing diplonemids

Horizontal gene transfer (HGT) is one of the fundamental processes in the evolution of prokaryotic genomes, while its importance in eukaryotes is still debated. Some of the hallmark eukaryotic organelles, such as mitochondria and chloroplasts, are of an ancient endosymbiotic origin. The process of acquiring (and losing) new endosymbionts is dynamic and still ongoing in many lineages. Endosymbiotic gene transfer (EGT) between symbionts and their hosts has been considered as one of the major sources of overall HGTs in eukaryotes. Thanks to recent advances in genomics and microscopy, more and more endosymbionts are discovered in protists offering suitable models for the study of EGT and its impact on the host. Recently, the presence of holosporacean and chlamydiacean symbionts in the novel strains of marine euglenozoan flagellates of genus Rhynchopus has been discovered. Here, we present an analysis of the genomes and transcriptomes of five Rhynchopus strains and examine the extent of EGT/HGT and the role of endosymbiosis in shaping the nuclear genome of symbiont-bearing hosts. Our results have shown that there is no evidence of a recent EGT from either Holosporales or Chlamydiales symbionts. The absence of such transfers suggests that EGT or at least a stable retention of EGT genes is not a requisite for a successful endosymbiosis. Furthermore, our results show striking differences between patterns of detected HGTs among the Rhynchopus lineages pointing to a dynamic and largely neutral evolution of horizontally-acquired genes.

evolutionary biology↗

Complexome profile of mitochondrial complexes in the myzozoan parasite of oysters, Perkinsus marinus

Mitochondrial complexes, such as the mitochondrial electron transport chain (mETC), the F1Fo-ATP synthase and the mitochondrial ribosome, are centrally important for mitochondrial function. Recently, an unexpected diversity in the composition of these complexes across unicellular eukaryotic lineages, with the apicomplexan parasites featuring prominently, has been revealed. However, whether the observed enlarged and divergent mitochondrial complexes are conserved across the wider Myzozoan lineage has not been investigated. Here, using a complexome profiling proteomic approach, we have characterised the composition of the mitochondrial complexes of the myzozoan parasite of oysters, Perkinsus marinus. We show that it shares with Plasmodium and Toxoplasma huge ATP synthase and mETC complexes. Moreover, the Perkinsus mitoribosome possesses many of the divergent features recently discovered in apicomplexans, such as an expanded subunit repertoire and the inclusion of RAP-domain and ApiAP2-like proteins as ribosomal proteins. Our study reveals the divergent subunit composition of mitochondrial complexes is an ancestral and highly conserved feature of Myzozoa.

microbiology↗

The first RNA viruses detected in a trypanosome: novel narnaviruses and a leishmaniavirus in the gecko parasite Trypanosoma platydactyli

BackgroundTrypanosomatids are parasitic flagellates best known for human pathogens causing sleeping sickness, Chagas disease, and leishmaniasis. RNA viruses infecting these protists have recently gained attention for their role in disease severity. While numerous such viruses have been described in Leishmania and several other trypanosomatid genera, none have previously been documented in the iconic genus Trypanosoma. ResultsWe report the first discovery and molecular characterization of RNA viruses in trypanosomes, identifying a leishmaniavirus and two narnaviruses in a single strain of Trypanosoma platydactyli, a parasite of the common wall gecko. The leishmaniavirus genome revealed a conserved organization, including a putative ribosomal frameshift site and a hairpin-like secondary structure typical of the genus. Phylogenetic inference indicates that it is closely related to leishmaniaviruses from Old World Leishmania spp., consistent with shared vector ecology. The two narnaviruses have distinct origins, although both cluster with viruses of other trypanosomatids, suggesting historical exchanges among co-infecting parasites. ConclusionsOur study expands both the known diversity of RNA viruses in trypanosomatids and the range of trypanosomatid genera that host these viruses, providing guidance for future screening. We suggest that vector ecology--particularly feeding behavior--may influence viral acquisition by trypanosomes, explaining the previous absence of viral reports from intensively studied trypanosomes of medical relevance vectored by tsetse flies or kissing bugs. Therefore, overlooked species transmitted by Nematocera represent promising candidates for future viral discovery. This concept extends beyond trypanosomatids, providing a general framework for understanding the conditions that permit viral host switching by viruses among microeukaryotes.

microbiology↗

Subcellular proteomics of Paradiplonema papillatum reveals digestive capacity of the cell membrane and the plasticity of peroxisomes across euglenozoans

Diplonemids are among the most diverse and abundant protists in the deep ocean, have extremely complex and ancient cellular systems, and exhibit unique metabolic capacities. Despite this, we know very little about this major group of eukaryotes. To establish a model organism for comprehensive investigation, we performed subcellular proteomics on Paradiplonema papillatum and localized 4,870 proteins to 22 cellular compartments. We additionally confirmed the predicted location of several proteins by epitope tagging and fluorescence microscopy. To probe the metabolic capacities of P. papillatum, we explored the proteins predicted to the cell membrane compartment in our subcellular proteomics dataset. Our data revealed an accumulation of many carbohydrate active enzymes (CAZymes). Our predictions suggest that these CAZymes are exposed to extracellular space, supporting proposals that diplonemids may specialize in breaking down carbohydrates in plant and algal cell walls. Further exploration of carbohydrate metabolism revealed an evolutionary divergence in the function of glycosomes (modified peroxisomes) in diplonemids versus kinetoplastids. Our subcellular proteome provides a resource for future investigations into the unique cell biology of diplonemids.

cell biology↗

Discovery of unique mitotic mechanisms in Paradiplonema papillatum

Diplonemids are highly diverse and abundant marine plankton with significant ecological importance. However, little is known about their biology, even in the model diplonemid Paradiplonema papillatum whose genome sequence is available. Examining the subcellular localization of proteins using fluorescence microscopy is a powerful approach to infer their putative function. Here we report a plasmid-based method that enables YFP-tagging of a gene at the endogenous locus. By examining the localization of proteins whose homologs are involved in chromosome segregation in other eukaryotes, we discovered several interesting features in mitotically dividing P. papillatum cells. Cohesin is enriched on condensed interphase chromatin. During mitosis, chromosomes organize into two rings (termed metaphase rings herein) that surround the elongating nucleolus. Homologs of chromosomal passenger complex components (INCENP, two Aurora kinases, and KIN-A), a CLK1 kinase, spindle checkpoint protein Mad1, and microtubule regulator XMAP215 localize in between the two metaphase rings, suggesting that kinetochores may assemble in between them. We also found that a homolog of the meiotic chromosome axis protein SYCP2L1 is enriched in between metaphase rings during mitosis. These features have some resemblance to the bivalent bridge (also known as the modified synaptonemal complex), which is thought to mediate the linkage between homologous chromosomes using axis components in Bombyx mori female meiotic metaphase I. By representing the first molecular characterization of mitotic mechanisms in P. papillatum and raising a number of questions, this study forms the foundation for dissecting mitotic mechanisms in diplonemids.

cell biology↗

Probing mechanical selection in diverse eukaryotic genomes through accurate prediction of 3D DNA mechanics

Connections between the mechanical properties of DNA and biological functions have been speculative due to the lack of methods to measure or predict DNA mechanics at scale. Recently, a proxy for DNA mechanics, cyclizability, was measured by loop-seq and enabled genome-scale investigation of DNA mechanics. Here, we use this dataset to build a computational model predicting bias-corrected intrinsic cyclizability, with near-perfect accuracy, solely based on DNA sequence. Further, the model predicts intrinsic bending direction in 3D space. Using this tool, we aimed to probe mechanical selection - that is, the evolutionary selection of DNA sequence based on its mechanical properties - in diverse circumstances. First, we found that the intrinsic bend direction of DNA sequences correlated with the observed bending in known protein-DNA complex structures, suggesting that many proteins co-evolved with their DNA partners to capture DNA in its intrinsically preferred bent conformation. We then applied our model to large-scale yeast population genetics data and showed that centromere DNA element II, whose consensus sequence is unknown, leaving its sequence-specific role unclear, is under mechanical selection to increase the stability of inner-kinetochore structure and to facilitate centromeric histone recruitment. Finally, in silico evolution under strong mechanical selection discovered hallucinated sequences with cyclizability values so extreme that they required experimental validation, yet, found in nature in the densely packed mitochondrial(mt) DNA of Namystynia karyoxenos, an ocean-dwelling protist with extreme mitochondrial gene fragmentation. The need to transmit an extraordinarily large amount of mtDNA, estimated to be > 600 Mb, in combination with the absence of mtDNA compaction proteins may have pushed mechanical selection to the extreme. Similarly extreme DNA mechanics are observed in bird microchromosomes, although the functional consequence is not yet clear. The discovery of eccentric DNA mechanics in unrelated unicellular and multicellular eukaryotes suggests that we can predict extreme natural biology which can arise through strong selection. Our methods offer a way to study the biological functions of DNA mechanics in any genome and to engineer DNA sequences with desired mechanical properties.

biophysics↗

On the possibility of yet a third kinetochore system in the protist phylum Euglenozoa

Transmission of genetic material from one generation to the next is a fundamental feature of all living cells. In eukaryotes, a macromolecular complex called the kinetochore plays crucial roles during chromosome segregation by linking chromosomes to spindle microtubules. Little is known about this process in evolutionarily diverse protists. Within the supergroup Discoba, Euglenozoa forms a speciose group of unicellular flagellates - kinetoplastids, euglenids, and diplonemids. Kinetoplastids have an unconventional kinetochore system, while euglenids have subunits that are conserved amongst most eukaryotes. For diplonemids, a group of extremely diverse and abundant marine flagellates, it remains unclear what kind of kinetochores are present. Here, we employed deep homology detection protocols using profile-versus-profile Hidden Markov Model searches and AlphaFold-based structural comparisons to detect homologies that might have been previously missed. Interestingly, we still could not detect orthologs for most of the kinetoplastid nor canonical kinetochore subunits with few exceptions including a putative centromere-specific histone H3 variant (cenH3/CENP-A), the spindle checkpoint protein Mad2, the chromosomal passenger complex members Aurora and INCENP, and broadly conserved proteins like CLK kinase and the meiotic synaptonemal complex proteins SYCP2/3 that also function at kinetoplastid kinetochores. We examined the localization of five candidate kinetochore-associated proteins in the model diplonemid, Paradiplonema papillatum. PpCENP-A shows discrete dots in the nucleus, implying that it is likely a kinetochore component. PpMad2, PpCLKKKT10/19, PpSYCP2L1KKT17/18 and INCENP reside in the nucleus, but no clear kinetochore localization was observed. Altogether, these results point to the possibility that diplonemids evolved a hitherto unknown type of kinetochore system. IMPORTANCEA macromolecular assembly called the kinetochore is essential for the segregation of genetic material during eukaryotic cell division. Therefore, characterization of kinetochores across species is essential for understanding the mechanisms involved in this key process across the eukaryotic tree of life. In particular, little is known about kinetochores in divergent protists such as Euglenozoa, a group of unicellular flagellates that includes kinetoplastids, euglenids, and diplonemids, the latter being a highly diverse and abundant component of marine plankton. While kinetoplastids have an unconventional kinetochore system and euglenids have a canonical one similar to traditional model eukaryotes, preliminary searches detected neither unconventional nor canonical kinetochore components in diplonemids. Here, we employed state-of-the-art deep homology detection protocols but still could not detect orthologs for the bulk of kinetoplastid-specific nor canonical kinetochore proteins in diplonemids except for a putative centromere-specific histone H3 variant. Our results strongly suggest that diplonemids evolved kinetochores that do not resemble previously known ones.

cell biology↗

Characterization of novel and essential kinetoplast-associated proteins in Trypanosoma brucei

The kinetoplast is one of the defining features of kinetoplastid protists and represents a unique concentration of mitochondrial DNA. This subcellular structure is a highly complex assembly of thousands of mutually catenated, circular DNA molecules as well as up to one hundred dedicated proteins. These components work in tandem to replicate and segregate the mitochondrial genome during cellular division, additionally coordinating with the basal body and flagellum through the tripartite attachment complex (TAC) superstructure. Here, we screened the MitoTag localization repository and identified a number of previously undescribed hypothetical proteins exhibiting putative signals within the kinetoplast of Trypanosoma brucei. Through endogenous tagging we verify their association with the kinetoplast or TAC. The essentiality for several of these kinetoplast proteins (KP) was assessed by RNAi knock-downs, revealing that the newly characterized KP56, KP84 and KP86 are indispensable for growth of the procyclic stage. Additionally, KP37, KP56, and KP84 displayed alterations in the abundance of maxicircles or minicircles, while the depletion of KP84 and KP86 resulted in cell cycle alternations. Pulldown assays using the endogenously V5-tagged cell lines identified novel interactors, which were additionally subjected to endogenous tagging for subcellular localization, revealing two additional proteins (KP45 and KP66) with dual localization to the kinetoplast and throughout the mitochondrial lumen. This work represents the most extensive identification of novel KPs to date and provides a methodological pipeline for the characterization of remaining KPs to further understand this intricate molecular structure.

cell biology↗

A novel nabelschnur protein regulates segregation of the kinetoplast DNA in Trypanosoma brucei

The kinetoplast DNA (kDNA), a distinctive arrangement of mitochondrial DNA found in trypanosomatid protists, comprises a concatenated network of minicircles and maxicircles that undergo division and segregation once during each cell cycle. Despite the identification and characterization of numerous proteins involved in kDNA maintenance and replication, its segregation and the formation of the nabelschnur remain poorly understood on a molecular level. This enigmatic filamentous structure, transiently appearing in Trypanosoma brucei, connects the daughter kDNA networks prior to their complete segregation. Here, we characterize TbNAB70, a high mobility group box-like protein localized exclusively to the nabelschnur and the kDNA disc. Our findings demonstrate that TbNAB70 is critical for the segregation, but not replication, of kDNA, a so far unprecedented phenotype. Furthermore, structural predictions suggest that this protein holds the capacity to bind to kDNA illuminating the exact molecular mechanisms of segregation involved. Thus, we propose that TbNAB70 plays a pivotal role in the faithful and efficient segregation of the daughter kDNA networks.

molecular biology↗

Ribosomal A-site interactions with near-cognate tRNAs drive stop codon readthrough

tRNAs serve as a dictionary for the ribosome translating the genetic message from mRNA into a polypeptide chain. Besides this canonical role, tRNAs are involved in other processes like programmed stop codon readthrough (SC-RT). There, tRNAs with near-cognate anticodons to stop codons must outcompete release factors and incorporate into the ribosomal decoding center to prevent termination and allow translation to continue. However, not all near-cognate tRNAs promote efficient SC-RT. Here, we demonstrate that those that do, establish critical contacts between their anticodon stem (AS) and ribosomal proteins Rps30/eS30 and Rps25/eS25 forming the decoding site. Unexpectedly, the length and well-defined nature of the AS determines the strength of these contacts, which is reflected in organisms with reassigned stop codons. These findings open a new direction in tRNA biology that should facilitate the design of artificial tRNAs with specifically altered decoding abilities.

molecular biology↗

Functional differentiation of Sec13 paralogs in the euglenozoan protists

The {beta}-propeller protein Sec13 plays roles in at least three distinct processes by virtue of being a component of the COPII endoplasmic reticulum export vesicle coat, the nuclear pore complex (NPC) and the Seh1-associated (SEA)/GATOR nutrient-sensing complex, suggesting that regulatory mechanisms coordinating these cellular activities may operate via Sec13. The NPC, COPII and SEA/GATOR are all ancient features of eukaryotic cells. In the vast majority of eukaryotes, a single Sec13 gene is present, but here we report that the Euglenozoa, a lineage encompassing the diplonemid, kinetoplastid and euglenid protists contain two Sec13 paralogs. Furthermore, based on protein interactions and localization studies we show that Sec13 functions in diplonemids are divided between the Sec13a and Sec13b paralogs. Specifically, Sec13a interacts with COPII and the NPC, but Sec13b interacts with Sec16 and components of the SEA/GATOR complex. We infer that euglenozoan Sec13a is responsible for NPC functions and canonical anterograde transport activities while Sec13b acts within nutrient and autophagy-related pathways, indicating a fundamentally distinct organization of coatomer complexes in the euglenozoan flagellates.

evolutionary biology↗

Massive accumulation of strontium and barium in diplonemid protists

Barium (Ba) and strontium (Sr) are often used as proxies for the reconstruction of past marine productivity and global climate. The ability to accumulate Ba2+ and Sr2+ in the form of crystals is rare among eukaryotes. Here we report that unicellular heterotrophs called diplonemids (Euglenozoa), one of the most abundant groups of marine planktonic protists, accumulate conspicuous amounts of these trace elements in the form of intracellular barite (BaSO4) and celestite (SrSO4) crystals, in concentrations greater than in other known Ba/Sr-accumulating organisms. Moreover, these flagellates can uptake Sr2+ exclusively or together with Ba2+ and form (Ba,Sr)SO4. One species, Namystinia karyoxenos, is naturally capable of intracellular accumulation of Ba2+ and Sr2+ 42,000 and 10,000 times relative to the surrounding medium. Altering the amounts of both elements in the medium resulted in corresponding changes in the quantity and composition of crystals. Planktonic copepods fed with diplonemids produce celestite-rich fecal pellets, which facilitate deposition of these minerals on the seafloor. We propose that diplonemids, which emerged during the Neoproterozoic era, qualify as impactful players of Ba2+/Sr2+ cycling in the ocean that possibly contributed to sedimentary rock formation over long geological periods.

microbiology↗

The development of intracytoplasmic membranes in alphaproteobacteria involves the conserved mitochondrial crista-developing Mic60 protein

Mitochondrial cristae expand the surface area of respiratory membranes and ultimately allow for the evolutionary scaling of respiration with cell volume across eukaryotes. The discovery of Mic60 homologs among alphaproteobacteria, the closest extant relatives of mitochondria, suggested that cristae might have evolved from bacterial intracytoplasmic membranes (ICMs). Here, we investigated the predicted structure and function of alphaproteobacterial Mic60, and a protein encoded by an adjacent gene Orf52, in two distantly related purple alphaproteobacteria, Rhodobacter sphaeroides and Rhodopseudomonas palustris. In addition, we assessed the potential physical interactors of Mic60 and Orf52 in R. sphaeroides. We show that the three -helices of mitochondrial Mic60s mitofilin domain, as well as its adjacent membrane-binding amphipathic helix, are present in alphaproteobacterial Mic60. The disruption of Mic60 and Orf52 caused photoheterotrophic growth defects, which are most severe under low light conditions, and both their disruption and overexpression led to enlarged ICMs in both studied alphaproteobacteria. We also found that alphaproteobacterial Mic60 physically interacts with BamA, the homolog of Sam50, one of the main physical interactors of eukaryotic Mic60. This interaction, responsible for making contact sites at mitochondrial envelopes, has been conserved in modern alphaproteobacteria despite more than a billion years of evolutionary divergence. Our results suggest a role for Mic60 in photosynthetic ICM development and contact site formation at alphaproteobacterial envelopes. Overall, we provide support for the hypothesis that mitochondrial cristae evolved from alphaproteobacterial ICMs, and therefore have improved our understanding of the nature of the mitochondrial ancestor.

evolutionary biology↗

Trypanosome morphogenesis involves recruitment of a pleckstrin homology domain protein by an orphan kinesin to the microtubule quartet

Kinesins are motor proteins found in all eukaryotic lineages that move along microtubules to mediate cellular processes such as mitosis and intracellular transport. In trypanosomatids, the kinesin superfamily has undergone a prominent expansion, resulting in one of the most diverse kinesin repertoires that includes the two kinetoplastid-restricted families X1 and X2. Here, we characterize in Trypanosoma brucei TbKifX2A, an orphaned X2 kinesin. TbKifX2A tightly interacts with TbPH1, a kinesin-like protein with a likely inactive motor domain, a rarely reported occurrence. Both TbKifX2A and TbPH1 localize to the microtubule quartet (MtQ), a characteristic but poorly understood cytoskeletal structure that wraps around the flagellar pocket as it extends to the cell body anterior. The proximal proteome of TbPH1 revealed two other interacting proteins, the flagellar pocket protein FP45 and intriguingly another X2 kinesin, TbKifX2C. Simultaneous ablation of TbKifX2A/TbPH1 results in the depletion of FP45 and TbKifX2C and also an expansion of the flagellar pocket, among other morphological defects. TbKifX2A is the first motor protein to be localized to the MtQ. The observation that TbKifX2C also associates with the MtQ suggests that the X2 kinesin family may have co-evolved with the MtQ, both kinetoplastid-specific traits.

molecular biology↗

Genetic tool development in marine protists: Emerging model organisms for experimental cell biology

Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and advancement of tools for 8 other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways.

ecology↗

Highly reduced genomes of protist endosymbionts show evolutionary convergence

Genome evolution in bacterial endosymbionts is notoriously extreme: the combined effects of strong genetic drift and unique selective pressures result in highly reduced genomes with distinctive adaptations to hosts [1-4]. These processes are mostly known from animal endosymbionts, where nutritional endosymbioses represent the best-studied systems. However, eukaryotic microbes, or protists, also harbor diverse bacterial endosymbionts, but their genome reduction and functional relationships with their more diverse hosts are largely unexplored [5-7]. We sequenced the genomes of four bacterial endosymbionts from three species of diplonemids, poorly-studied but abundant and diverse heterotrophic protists [8-10]. The endosymbionts come from two intracellular families from different orders, Rickettsiaceae and Holosporaceae, that have invaded diplonemids multiple times, and their genomes have converged on an extremely small size (605-632 kbp), similar gene content (e.g., metabolite transporters and secretion systems), and reduced metabolic potential (e.g., loss of energy metabolism). These characteristics are generally found in both families, but the diplonemid endosymbionts have evolved greater extremes in parallel. Their modified type VI secretion systems are likely involved in the manipulation of host metabolism (e.g., interactions with host mitochondria) or defense against bacterial infections, although their similar effector/immunity proteins may also allow for co-occurring Holosporaceae species in one diplonemid host. Finally, modified cellular machinery like ATP synthase without oxidative phosphorylation and reduced flagella present in both diplonemid endosymbionts and nutritional animal endosymbionts indicates that intracellular mechanisms have converged in bacterial endosymbionts with various functions and from different eukaryotic hosts across the tree of life.

evolutionary biology↗