Eukaryotic Origin Research Articles

The Proterozoic macrofossil Tawuia as a coenocytic eukaryote and a possible macroalga

Proterozoic carbonaceous macrofossils are key to understand the evolution of early eukaryotes and how they acquired macroscopic sizes. However, phylogenetic interpretation of these macrofossils is challenging because most of them are morphologically simple and lack phylogenetically diagnostic features. Among Proterozoic carbonaceous macrofossils, the tomaculate fossil Tawuia is relatively rich in characters and is common in Proterozoic successions. However, its phylogenetic interpretation is uncertain and Tawuia has been variously interpreted as a bacterium or an alga (photosynthetic eukaryote), in part because certain aspects of its morphologies have not been consistently documented, hampering their functional interpretations. Here we report exceptionally preserved Tawuia specimens from the Tonian Liulaobei and Shiwangzhuang formations in North China. These fossils show that Tawuia is a tomaculate organism, sometimes with an axial stripe, terminal discoidal structures, and transverse constrictions, features that were previously documented but variously interpreted. We infer that Tawuia was a coenocytic organism with recalcitrant cell walls, asexual reproduction through asymmetric constrictions, and possible abscission structures. These features indicate that Tawuia is probably a coenocytic eukaryote, and possibly a macroalga, although more evidence is needed to confirm these interpretations, particularly the photosynthetic nature of Tawuia. A literature review reveals that the key features described here occur not only in many Tonian Tawuia specimens, but also in some possible Mesoproterozoic Tawuia fossils as much as ca. 1.6 Ga old. The new data and interpretations of Tawuia presented here provide important constraints on the origin and early evolution of coenocytic eukaryotes, primary plastids, and macroalgae, particularly if the Mesoproterozoic Tawuia fossils are confirmed as macroalgae.

Evidence for a Syncytial Origin of Eukaryotes from Ancestral State Reconstruction.

Modern accounts of eukaryogenesis entail an endosymbiotic encounter between an archaeal host and a proteobacterial endosymbiont, with subsequent evolution giving rise to a unicell possessing a single nucleus and mitochondria. The mononucleate state of the last eukaryotic common ancestor (LECA) is seldom, if ever, questioned, even though cells harboring multiple (syncytia, coenocytes, and polykaryons) are surprisingly common across eukaryotic supergroups. Here, we present a survey of multinucleated forms. Ancestral character state reconstruction for representatives of 106 eukaryotic taxa using 16 different possible roots and supergroup sister relationships, indicate that LECA, in addition to being mitochondriate, sexual, and meiotic, was multinucleate. LECA exhibited closed mitosis, which is the rule for modern syncytial forms, shedding light on the mechanics of its chromosome segregation. A simple mathematical model shows that within LECA’s multinucleate cytosol, relationships among mitochondria and nuclei were neither one-to-one, nor one-to-many, but many-to-many, placing mitonuclear interactions and cytonuclear compatibility at the evolutionary base of eukaryotic cell origin. Within a syncytium, individual nuclei and individual mitochondria function as the initial lower-level evolutionary units of selection, as opposed to individual cells, during eukaryogenesis. Nuclei within a syncytium rescue each other’s lethal mutations, thereby postponing selection for viable nuclei and cytonuclear compatibility to the generation of spores, buffering transitional bottlenecks at eukaryogenesis. The prokaryote-to-eukaryote transition is traditionally thought to have left no intermediates, yet if eukaryogenesis proceeded via a syncytial common ancestor, intermediate forms have persisted to the present throughout the eukaryotic tree as syncytia but have so far gone unrecognized.

Expanded diversity of Asgard archaea and their relationships with eukaryotes.

Asgard is a recently discovered superphylum of archaea that appears to include the closest archaeal relatives of eukaryotes1-5. Debate continues as to whether the archaeal ancestor of eukaryotes belongs within the Asgard superphylum or whether this ancestor is a sister group to all other archaea (that is, a two-domain versus a three-domain tree of life)6-8. Here we present a comparative analysis of 162complete or nearly complete genomes of Asgard archaea, including 75metagenome-assembled genomes that-to our knowledge-have not previously been reported. Our results substantially expand the phylogenetic diversity of Asgard and lead us to propose six additional phyla that include a deep branch that we have provisionally named Wukongarchaeota. Our phylogenomic analysis does not resolve unequivocally the evolutionary relationship between eukaryotes and Asgard archaea, but instead-depending on the choice of species and conserved genes used to build the phylogeny-supports either the origin of eukaryotes from within Asgard (as a sister group to the expanded Heimdallarchaeota-Wukongarchaeota branch) or a deeper branch for the eukaryote ancestor within archaea. Our comprehensive protein domain analysis using the 162Asgard genomes results in a major expansion of the set of eukaryotic signature proteins. The Asgard eukaryotic signature proteins show variable phyletic distributions and domain architectures, which is suggestive of dynamic evolution through horizontal gene transfer, gene loss, gene duplication and domain shuffling. The phylogenomics of the Asgard archaea points to the accumulation of the components of the mobile archaeal 'eukaryome' in the archaeal ancestor of eukaryotes (within or outside Asgard) through extensive horizontal gene transfer.

Substrate Tunnel Engineering Aided by X-ray Crystallography and Functional Dynamics Swaps the Function of MIO-Enzymes

The enzyme family harboring the post-translationally formed 5-methylene-3,5-dihydro-4H-imidazol-4-one (MIO) catalytic residue comprises both aromatic amino acid ammonia-lyases (ALs) and 2,3-aminomutases (AMs). The structural origin of the different functions and the role of the inner loop region in substrate binding are not fully understood. Here, we provide the three-dimensional structures for Petroselinum crispum phenylalanine AL (PcPAL) with fully resolved inner loops in a catalytically competent conformation. Using molecular modeling, we demonstrate that in both ALs and AMs of eukaryotic origin, just a small opening of the inner loop is sufficient for ligand egress. Furthermore, we show that ligand-binding tunnels are analogous to eukaryotic MIO-enzymes and that the critical initial part of these tunnels is present across the whole enzyme family. Engineering of these binding tunnels converts an (R)-AM to a highly selective (S)-β-AL thus establishing a nonclassified enzyme function.

Gene Duplications Trace Mitochondria to the Onset of Eukaryote Complexity.

The last eukaryote common ancestor (LECA) possessed mitochondria and all key traits that make eukaryotic cells more complex than their prokaryotic ancestors, yet the timing of mitochondrial acquisition and the role of mitochondria in the origin of eukaryote complexity remain debated. Here, we report evidence from gene duplications in LECA indicating an early origin of mitochondria. Among 163,545 duplications in 24,571 gene trees spanning 150 sequenced eukaryotic genomes, we identify 713 gene duplication events that occurred in LECA. LECA’s bacterial-derived genes include numerous mitochondrial functions and were duplicated significantly more often than archaeal-derived and eukaryote-specific genes. The surplus of bacterial-derived duplications in LECA most likely reflects the serial copying of genes from the mitochondrial endosymbiont to the archaeal host’s chromosomes. Clustering, phylogenies and likelihood ratio tests for 22.4 million genes from 5,655 prokaryotic and 150 eukaryotic genomes reveal no evidence for lineage-specific gene acquisitions in eukaryotes, except from the plastid in the plant lineage. That finding, and the functions of bacterial genes duplicated in LECA, suggests that the bacterial genes in eukaryotes are acquisitions from the mitochondrion, followed by vertical gene evolution and differential loss across eukaryotic lineages, flanked by concomitant lateral gene transfer among prokaryotes. Overall, the data indicate that recurrent gene transfer via the copying of genes from a resident mitochondrial endosymbiont to archaeal host chromosomes preceded the onset of eukaryotic cellular complexity, favoring mitochondria-early over mitochondria-late hypotheses for eukaryote origin.

Prokaryotic symbiotic consortia and the origin of nucleated cells: A critical review of Lynn Margulis hypothesis

The publication in the late 1960s of Lynn Margulis endosymbiotic proposal is a scientific milestone that brought to the fore of evolutionary discussions the issue of the origin of nucleated cells. Although it is true that the times were ripe, the timely publication of Lynn Margulis' original paper was the product of an intellectually bold 29-years old scientist, who based on the critical analysis of the available scientific information produced an all-encompassing, sophisticated narrative scheme on the origin of eukaryotic cells as a result of the evolution of prokaryotic consortia and, in bold intellectual stroke, put it all in the context of planetary evolution. A critical historical reassessment of her original proposal demonstrates that her hypothesis was not a simple archival outline of past schemes, but a renewed historical narrative of prokaryotic evolution and the role of endosymbiosis in the origin of eukaryotes. Although it is now accepted that the closest bacterial relatives of mitochondria and plastids are α-proteobacteria and cyanobacteria, respectively, comparative genomics demonstrates the mosaic character of the organelle genomes. The available evidence has completely refuted Margulis’ proposal of an exogenous origin for eukaryotic flagella, the (9 + 2) basal bodies, and centromeres, but we discuss in detail the reasons that led her to devote considerable efforts to argue for a symbiotic origin of the eukaryotic motility. An analysis of the arguments successfully employed by Margulis in her persuasive advocacy of endosymbiosis, combined with the discussions of her flaws and the scientific atmosphere during the period in which she formulated her proposals, are critical for a proper appraisal of the historical conditions that shaped her theory and its acceptance.

Proline Rich Peptides of Neurohypophysial Origin: Related Peptides and Possible Functions

Proline rich polypeptide 1 (PRP-1) was isolated from the neurosecretory granules of bovine neurohypophysis and thoroughly characterized by Professor A. Galoyan and coauthors. BLAST analyses of PRP-1 with proteins of protein data bank indicated, that it is the C-terminus of third constituent of pre-pro-arginine vasopressin – copeptin, for all studied mammal species, as showed Galoyan and coauthors for bovine case. Also, BLAST analyses indicated, that the amino acid motive of PRP-1 is conservative in the course of the evolution, and occurred among enzymes of prokaryotic and eukaryotic origin Obtained to date data indicate multiple functionalities of this polypeptide, through to its diverse modulatory actions on set of different receptors. Using the AutoDock vina software it was shown, that PRP-1 and its homologues strongly interact with receptors, such as: mouse phosphorylated mitogen activated protein kinase 14, mouse toll like receptor 4, human Ca-sensing receptor, human epithelial growth factor receptor and human superoxide dismutase 1. As these receptors play important role in multiple cell signaling processes, the importance of their ligand peptides and homologues is obvious. But it should be stated, that, through docking experiments although are very informative, they should be considered as supplementary to real physical experiments, which should be carried out with the same receptors.

Probing the effects of double mutations on the versatile protein ubiquitin in Saccharomyces cerevisiae

Ubiquitin is an indispensable protein of eukaryotic origin with an extraordinarily high degree of sequence conservation. It is used to tag proteins post-translationally and the process of ubiquitination regulates the activity of the modified proteins or drives them for degradation. Double mutations produce varied effects in proteins, depending on the structural relationship of the mutated residues, their role in the overall structure and functions of a protein. Six double mutants derived from the ubiquitin mutant UbEP42, namely S20F-A46S, S20F-L50P, S20F-I61T, A46S-L50P, A46S-I61T, and L50P-I61T, have been studied here to understand how they influence the ubiquitination related functions, by analysing their growth and viability, Cdc28 levels, K-48 linked polyubiquitination, UFD pathway, lysosomal degradation, endosomal sorting, survival under heat, and antibiotic stresses. The double mutation L50P-I61T is the most detrimental, followed by S20F-I61T and A46S-I61T. The double mutations studied here, in general, make cells more sensitive than the wild type to one or the other stress. However, the excessive negative effects of L50P and I61T are compensated under certain conditions by S20F and A46S mutations. The competitive inhibition produced by these substitutions could be used to manage certain ubiquitination associated diseases.

MTBP phosphorylation controls DNA replication origin firing

Faithful genome duplication requires regulation of origin firing to determine loci, timing and efficiency of replisome generation. Established kinase targets for eukaryotic origin firing regulation are the Mcm2-7 helicase, Sld3/Treslin/TICRR and Sld2/RecQL4. We report that metazoan Sld7, MTBP (Mdm2 binding protein), is targeted by at least three kinase pathways. MTBP was phosphorylated at CDK consensus sites by cell cycle cyclin-dependent kinases (CDK) and Cdk8/19-cyclin C. Phospho-mimetic MTBP CDK site mutants, but not non-phosphorylatable mutants, promoted origin firing in human cells. MTBP was also phosphorylated at DNA damage checkpoint kinase consensus sites. Phospho-mimetic mutations at these sites inhibited MTBP’s origin firing capability. Whilst expressing a non-phospho MTBP mutant was insufficient to relieve the suppression of origin firing upon DNA damage, the mutant induced a genome-wide increase of origin firing in unperturbed cells. Our work establishes MTBP as a regulation platform of metazoan origin firing.

Coevolutionary and Phylogenetic Analysis of Mimiviral Replication Machinery Suggest the Cellular Origin of Mimiviruses.

Mimivirus is one of the most complex and largest viruses known. The origin and evolution of Mimivirus and other giant viruses have been a subject of intense study in the last two decades. The two prevailing hypotheses on the origin of Mimivirus and other viruses are the reduction hypothesis, which posits that viruses emerged from modern unicellular organisms; whereas the virus-first hypothesis proposes viruses as relics of precellular forms of life. In this study, to gain insights into the origin of Mimivirus, we have carried out extensive phylogenetic, correlation, and multidimensional scaling analyses of the putative proteins involved in the replication of its 1.2-Mb large genome. Correlation analysis and multidimensional scaling methods were validated using bacteriophage, bacteria, archaea, and eukaryotic replication proteins before applying to Mimivirus. We show that a large fraction of mimiviral replication proteins, including polymerase B, clamp, and clamp loaders are of eukaryotic origin and are coevolving. Although phylogenetic analysis places some components along the lineages of phage and bacteria, we show that all the replication-related genes have been homogenized and are under purifying selection. Collectively our analysis supports the idea that Mimivirus originated from a complex cellular ancestor. We hypothesize that Mimivirus has largely retained complex replication machinery reminiscent of its progenitor while losing most of the other genes related to processes such as metabolism and translation.

Opinion: Genetic Conflict With Mobile Elements Drives Eukaryotic Genome Evolution, and Perhaps Also Eukaryogenesis.

Through analyses of diverse microeukaryotes, we have previously argued that eukaryotic genomes are dynamic systems that rely on epigenetic mechanisms to distinguish germline (i.e., DNA to be inherited) from soma (i.e., DNA that undergoes polyploidization, genome rearrangement, etc.), even in the context of a single nucleus. Here, we extend these arguments by including two well-documented observations: (1) eukaryotic genomes interact frequently with mobile genetic elements (MGEs) like viruses and transposable elements (TEs), creating genetic conflict, and (2) epigenetic mechanisms regulate MGEs. Synthesis of these ideas leads to the hypothesis that genetic conflict with MGEs contributed to the evolution of a dynamic eukaryotic genome in the last eukaryotic common ancestor (LECA), and may have contributed to eukaryogenesis (i.e., may have been a driver in the evolution of FECA, the first eukaryotic common ancestor). Sex (i.e., meiosis) may have evolved within the context of the development of germline-soma distinctions in LECA, as this process resets the germline genome by regulating/eliminating somatic (i.e., polyploid, rearranged) genetic material. Our synthesis of these ideas expands on hypotheses of the origin of eukaryotes by integrating the roles of MGEs and epigenetics.

Experimental taphonomy of organelles and the fossil record of early eukaryote evolution.

The timing of origin of eukaryotes and the sequence of eukaryogenesis are poorly constrained because their fossil record is difficult to interpret. Claims of fossilized organelles have been discounted on the unsubstantiated perception that they decay too quickly for fossilization. We experimentally characterized the pattern and time scale of decay of nuclei, chloroplasts, and pyrenoids in red and green algae, demonstrating that they persist for many weeks postmortem as physical substrates available for preservation, a time scale consistent with known mechanisms of fossilization. Chloroplasts exhibit greater decay resistance than nuclei; pyrenoids are unlikely to be preserved, but their presence could be inferred from spaces within fossil chloroplasts. Our results are compatible with differential organelle preservation in seed plants. Claims of fossilized organelles in Proterozoic fossils can no longer be dismissed on grounds of plausibility, prompting reinterpretation of the early eukaryotic fossil record and the prospect of a fossil record of eukaryogenesis.

Toward simulating the origin of eukaryotes in an artificial cell

Toward simulating the origin of eukaryotes in an artificial cell

Recombinant production of membrane proteins in yeast.

Biochemical pathways are compartmentalized in living cells. This permits each cell to maintain chemical compositions that differ between the cytosol, intracellular organelles and the external environment. Achieving this requires each compartment to be very selective in what is allowed to enter and leave. Nature has solved this by surrounding each cell and each organelle with a virtually solute impermeable lipid membrane, embedded with integral membrane proteins that mediate strictly controlled trans-membrane movement of matter and information. Access to pure and active integral membrane proteins is therefore required to comprehend membrane biology, ultimately through high-resolution structures of the membrane proteome and, therefore, also for our understanding of physiology. Unfortunately, apart from a few exceptions, membrane proteins cannot be purified from native tissue but need to be produced recombinantly, which is eminently challenging. This chapter shows how we have engineered yeast to provide high levels of prime quality membrane proteins of prokaryotic, archaeal or eukaryotic origin for structural biology.

Importance of Prokaryotes for the Origin of Eukaryotes and the Global Environment at 2.4-2.0 Ga

Importance of Prokaryotes for the Origin of Eukaryotes and the Global Environment at 2.4-2.0 Ga

S-adenosyl-l-homocysteine Hydrolase: A Structural Perspective on the Enzyme with Two Rossmann-Fold Domains.

S-adenosyl-l-homocysteine hydrolase (SAHase) is a major regulator of cellular methylation reactions that occur in eukaryotic and prokaryotic organisms. SAHase activity is also a significant source of l-homocysteine and adenosine, two compounds involved in numerous vital, as well as pathological processes. Therefore, apart from cellular methylation, the enzyme may also influence other processes important for the physiology of particular organisms. Herein, presented is the structural characterization and comparison of SAHases of eukaryotic and prokaryotic origin, with an emphasis on the two principal domains of SAHase subunit based on the Rossmann motif. The first domain is involved in the binding of a substrate, e.g., S-adenosyl-l-homocysteine or adenosine and the second domain binds the NAD+ cofactor. Despite their structural similarity, the molecular interactions between an adenosine-based ligand molecule and macromolecular environment are different in each domain. As a consequence, significant differences in the conformation of d-ribofuranose rings of nucleoside and nucleotide ligands, especially those attached to adenosine moiety, are observed. On the other hand, the chemical nature of adenine ring recognition, as well as an orientation of the adenine ring around the N-glycosidic bond are of high similarity for the ligands bound in the substrate- and cofactor-binding domains.

Modification and Expression of Beta-1,4-Endoglucanase encoding sequences of fungal origin in Escherichia coli BL21.

Lignocellulose is the main and most abundant component of biomass. Annually, 200 million tons are generated in the world. Colombia has a high production of lignocellulosic residues that can be used in many industrial processes such as bioethanol production, promoting the bioeconomy. The objective of the present work was to express lignocellulolytic enzymes of eukaryotic origin in Escherichia coli BL21 (DE3). Initially, endoglucanase eukaryotic genes were selected and modified using bioinformatics methods for their production in E. coli BL21 (DE3) and saccharification of pure cellulose substrates. The gene selected for its modification and expression was eglB from the fungus Aspergillus nidulans. Subsequently the enzyme integrity was tested by 3D modeling and molecular docking, as well as the conformation of its active site and its affinity for substrates of interest. Finally, cloning of the modified gene in plasmid pET151 TOPO was made and transformed in the strain E. coli BL21 (DE3) where several lignocellulose degradation tests were carried out using semiquantitative methods for the enzyme activity in carboxymethylcellulose. The presence of the three genes of interest within the plasmid pET151 TOPO and within the transformed cells of E. coli TOP10 and E. coli BL21 (DE3) was verified by colony PCRs performed. The presence of this gen was corroborated by sequencing. Expression of the modified endoglucanase enzyme was achieved in E. coli BL21 (DE3) expression cells, in soluble and functional form, demonstrated by the hydrolysis of the CMC substrate.

Ecological features and global distribution of Asgard archaea

Asgard is a newly proposed archaeal superphylum, which has been suggested to hold the key to decipher the origin of Eukaryotes. However, their ecology remains largely unknown. Here, we conducted a meta-analysis of publicly available Asgard-associated 16S rRNA gene fragments, and found that just three previously proposed clades (Lokiarchaeota, Thorarchaeota, and Asgard clade 4) are widely distributed, whereas the other seven clades (phylum or class level) are restricted to the sediment biosphere. Asgard archaea, especially Loki- and Thorarchaeota, seem to adapt to marine sediments, and water depth (the depth of the sediment below water surface) and salinity might be crucial factors for the proportion of these microorganisms as revealed by multivariate regression analyses. However, the abundance of Asgard archaea exhibited distinct environmental drivers at the clade-level; for instance, the proportion of Asgard clade 4 was higher in less saline environments (salinity <6.35 psu), while higher for Heimdallarchaeota-AAG and Asgard clade 2 in more saline environment (salinity ≥35 psu). Furthermore, co-occurrence analysis allowed us to find a significant non-random association of different Asgard clades with other groups (e.g., Lokiarchaeota with Deltaproteobacteria and Anaerolineae; Odinarchaeota with Bathyarchaeota), suggesting different interaction potentials among these clades. Overall, these findings reveal Asgard archaea as a ubiquitous group worldwide and provide initial insights into their ecological features on a global scale.

Green Synthesis of Metallic Nanoparticles and Their Potential Applications to Treat Cancer.

Nanoparticle synthesis using microorganisms and plants by green synthesis technology is biologically safe, cost-effective, and environment-friendly. Plants and microorganisms have established the power to devour and accumulate inorganic metal ions from their neighboring niche. The biological entities are known to synthesize nanoparticles both extra and intracellularly. The capability of a living system to utilize its intrinsic organic chemistry processes in remodeling inorganic metal ions into nanoparticles has opened up an undiscovered area of biochemical analysis. Nanotechnology in conjunction with biology gives rise to an advanced area of nanobiotechnology that involves living entities of both prokaryotic and eukaryotic origin, such as algae, cyanobacteria, actinomycetes, bacteria, viruses, yeasts, fungi, and plants. Every biological system varies in its capabilities to supply metallic nanoparticles. However, not all biological organisms can produce nanoparticles due to their enzymatic activities and intrinsic metabolic processes. Therefore, biological entities or their extracts are used for the green synthesis of metallic nanoparticles through bio-reduction of metallic particles leading to the synthesis of nanoparticles. These biosynthesized metallic nanoparticles have a range of unlimited pharmaceutical applications including delivery of drugs or genes, detection of pathogens or proteins, and tissue engineering. The effective delivery of drugs and tissue engineering through the use of nanotechnology exhibited vital contributions in translational research related to the pharmaceutical products and their applications. Collectively, this review covers the green synthesis of nanoparticles by using various biological systems as well as their applications.

Progress in elucidating the origin of eukaryotes

Knowledge of the origin of eukaryotes is key to broadening our understanding of the eukaryotic genome and the relationship among internal structures within a eukaryotic cell. Since the discovery of archaea in 1977 and the proposal of three-domain tree of life by the American microbiologist Carl Woese, the intimate relationship in evolution between eukaryotes and archaea has been demonstrated by considerable experiments and analyses. From the beginning of the 21st century, with the development of phylogenetic methods and the discovery of new archaeal phyla more related to eukaryotes, increasing evidence has shown that Eukarya and Archaea should be merged into one domain, leading to a two-domain tree of life. Nowadays, the Asgard superphylum discovered via metagenomic analysis is regarded as the closest prokaryotes to eukaryotes. Nevertheless, several key questions are still under debate, such as what the ancestors of the eukaryotes were and when mitochondria emerged. Here, we review the current research progress regarding the changes of the tree of life and the detailed eukaryotic evolutionary mechanism. We show that the recent findings have greatly improved our knowledge on the origin of eukaryotes, which will pave the way for future studies.