Most importantly, while these represent ancient proteins, none show the phylogenetic pattern that would be expected for a necessary stacking factor, illustrated in the Prediction row. To clarify the patterns of presence and absence in organisms with stacked and unstacked cisternae, only selected genomes are shown here. The full data are given in Additional file 2 : Figure S1 and Additional file 6 : Table. The first four columns (blue) show genes identified in organisms with unstacked Golgi, and closely related organisms with stacked Golgi, while remaining columns (red) indicate genes identified in representatives of taxonomic groups with stacked Golgi. Gray sectors indicate sequences identified using alternative methods (Additional file 2 : Figure S1). B Schematic showing the timing of gains and losses of the proteins across eukaryotic evolution. Note that, if a single member of the taxonomic group possesses precision an orthologue of the protein, it is inferred as present in that group. Relationships between eukaryotes are based on recent concatenated phylogenetic results 75, 101. To highlight losses in the Ascomycota, they are broken out to the exclusion of the paraphyletic remaining Fungi (denoted by the asterisk) The above observations suggest that the origin of both GM130 and golgin-45 predates the duplication that produced separate grasp55 and grasp65 paralogues, rather. Recent structural studies have elucidated the interaction between grasp65 and GM130 34, and between grasp55 and golgin-45 35, suggesting that these binding interactions involve specific residues near the c-terminus of GM130 and golgin-45 interacting with specific residues of grasp65 and grasp55, respectively.
Category: Golgi apparatus - wikimedia commons
A coulson plot of Metazoa-specific golgin complement. Note that, for this figure and Fig. 4, filled pie sectors represent the positive identification of at least one orthologue (paralogue numbers are not shown). Light blue sectors indicate instances where an orthologue was not found apivita in ciona intestinalis but was found in the genome of a closely related ascidian. This representation is based on data shown in Additional file 2 : Figure S1 and Additional file 6 : Table. B Schematic showing timing of gains and losses of metazoan golgin genes. Note that, here and for Fig. 4, gene duplications yielding expanded complements oily are not tracked and losses are only inferred when a factor was not identified in more than one representative of a taxonomic group Fig. 4 Pan-eukaryotic Golgi protein evolution. A coulson plot of Golgi proteins found outside the metazoa.
and golgin-45 (Fig. 3a ; Additional file 2 : Figure S1; Additional file 6 : Table S3) revealed no homologues outside of animals and their single-celled relatives (Holozoa). Consistent with previous efforts, our analysis did not identify the gm130 analogue bug1p as a homologue of GM130 in Saccharomyces based on sequence similarity. Homologues of grasp55 and grasp65 have been previously identified in diverse eukaryotes and functionally studied in organisms both with canonical stacked Golgi 32 and with unusual morphologies. Consistent with this result, and expanding upon it, we found that the duplication into grasp55 and grasp65 is a metazoan trait, predating the evolution of jawed fish (Additional file 7 : Figure S4 which means that all grasp proteins in other eukaryotes are pre-duplicates. Also consistent with previous analyses 24, 33, grasp was found across eukaryotes (Fig. 4a, additional file 2 : Figure S1, and Additional file 6 : Table S3) implying its presence in the leca. However, grasp was not identified in many cases, most prominently in Embryophyta as previously noted 33 and extended here to the entire clade of Archaeplastida plus Cryptophyta, as well as Rhizaria and Metamonada (Fig. 3 Metazoa-specific golgin evolution.
Ppt, golgi, powerPoint Presentation - id:3722092
Α tubulin antibody labelled the zeelandnet tubular conus around nuclei and network of fibers. Signal for pdi network is concentrated around multiple nuclei. Graphs show line scans for fluorescence intensities corresponding to the dotted lines in merged images. Scale bar, 5 μm The copi complex mediates traffic from the golgi to the er in eukaryotic cells, and therefore the er would be a likely location for the copi complex were a golgi not present. To ensure that this was not the case, we co-localized the copi-β with protein disulfide-isomerase (pdi a well-known er marker. This showed a pdi signal present in tubular structures close to nuclei as well as in numerous vesicles in the endoplasm, but little overlap with the copi-β signal (Fig. Furthermore, since hydrogenosomes, the mitochondria-derived organelles. Balamuthi, can also take the form of small discrete punctae 28, co-localization experiments were performed (Fig. 2, middle row) showing no overlap between copi-β and the hydrogenosomal marker malate dehydrogenase. Together, these informatics and microscopy results are most consistent with the presence of a cryptic vrouwen unstacked Golgi. Balamuthi, and validate the inclusion of genomic information from this organism in our subsequent searches. Evolution of the interacting Golgi structural proteins GM130, golgin-45, grasp55, and grasp65 to understand the distribution and evolution of proteins with putative roles in Golgi stacking, we performed comparative genomic searches to assess the taxonomic distribution of mammalian golgins, as well as other Golgi proteins.
Balamuthi cytosol, confirming expression of the protein and indicating a vesicular form of the organelle (Fig. We did not observe any association of Golgi with cytoskeletal structures of the microtubular conus around the cells multiple nuclei and microtubular fibers. Balamuthi with 10 nM, 100 nM, 1 μm, and 10 μm of Brefeldin A for 5 hours and subsequently analyzed the copi-β signal by sim. However, we did not observe any difference in comparison to non-treated cells (data not shown). Brefeldin a-insensitive versions of gbf1 (the Arfgef upon which Brefeldin acts) have been reported in other taxa, such as Arabidopsis 26 and Canis familiaris 27, and we suggest that this is likely the case here. Consistent with this hypothesis, the relevant amino acid residue for Brefeldin sensitivity in this protein (corresponding to M832 in Homo sapiens ) is not conserved. Balamuthi (for sequence see additional file 3 : Table S2). 2 Localization for. Structured illumination microscopy. Balamuthi labelled with antibodies against copi and pdi (top row, er structure mdh (middle row, hydrogenosomes and α tubulin (bottom row). The copi signal is observed in numerous vesicles scattered within the.
Camillo, golgi - himetop1 ; Additional file 3 : Table S2 including the neck soluble n -ethylmaleimide-sensitive fusion protein attachment protein receptor (snare) proteins Syn5, syn16, and Sec22, the retromer complex component Vps35, and the components of the multi-subunit tethering complexes that act at the golgi, cog and trappii. This list also includes the genes encoding the large subunits of the Adaptin 1, 3, and 4 complexes involved in transport from the trans- golgi network (tgn and the β-subunit of the coat protein complex I (copi) involved in intra-golgi transport and traffic from the. 1 diagram showing the golgi marker genes found. Balamuthi and their location in a generalized eukaryotic cell (see additional file 3 : Table S2 for further details). Notably, we identified proteins with roles in vesicle fusion and formation, transport to and from the golgi, and whose orthologues act at both the cis and trans faces of the organelle in other eukaryotes. Arrows indicate some membrane trafficking pathways that are reconstructed as likely present in the membrane trafficking system. Balamuthi golgi-like compartments. Balamuthi are dispersed and punctate to validate our genomic and informatics findings, we took a molecular cell biological approach. After further confirming the orthology of the copi-β orthologue. Balamuthi by phylogenetic analysis (Additional file 4 : Figure S2 a specific antibody was raised and validated (Additional file 5 : Figure S3 and used for immunofluorescence light microscopy. This showed localization to discrete punctate structures scattered throughout the.
Herein, we report an analysis of golgins and other Golgi structure-associated proteins across eukaryotes, using genomics, molecular cell biology, and bioinformatics techniques to address evolutionary cell biology of the golgi in eukaryotes. Results The genome of the golgi-less amoeba. Balamuthi encodes Golgi proteins Genome sequences exist for 11 microbial eukaryotes with evidence for the presence of a golgi, but presumably in an unstacked morphology. These organisms are spread throughout the diversity of eukaryotes (Additional file 2 : Figure S1 yet in the supergroup Amoebozoa only one genus, the parasitic Entamoeba, has an unstacked Golgi, which has been characterized to some extent. Balamuthi is berry a free-living anaerobic amoeba, related to Entamoeba, that lacks an identifiable stacked Golgi and that was at one time proposed to be lacking the organelle. To expand our sampling of eukaryotic genomes for this comparative analysis, particularly to increase taxon sampling in the Amoebozoa by adding a non-parasitic representative, we searched within the draft genome. Balamuthi (see methods ) for genes that might indicate the presence of a golgi. A set of Golgi marker genes has been previously established to have been present in the leca 24, and also as present in the genomes of organisms that lack golgi stacking 12, 16 19,. Previously seven such proteins were reported for. Balamuthi based on individual gene studies 12,. We were able to expand this list to a total of 22 proteins (Fig.
Golgi apparatus - wikipedia
Furthermore, several golgins, including GM130, are involved in connecting the golgi to the cytoskeleton 9,. Various additional proteins have also been suggested to be involved in Golgi structure and recept organization (Additional file 1 : Table S1). The integral role of golgins and other implicated structural proteins at the golgi makes their evolutionary histories essential to reconstructing both the nature of the golgi in the last eukaryotic common ancestor (leca) approximately.5 billion years ago 11, and to tracing the subsequent changes. While it has been inferred that the leca possessed a stacked Golgi 12, whether there are pan-eukaryotic proteins (e.g., golgins) that may have conserved roles in Golgi stacking remains unknown. Furthermore, the extent and details of golgin-mediated vesicle trafficking in the diversity of eukaryotes as compared with mammalian cells is also an open question. Intriguingly, while golgi stacking is observed in most organisms across eukaryotic diversity, there are a few lineages of microbial eukaryotes that lack stacked Golgi, as reviewed previously. In the absence of a morphologically recognizable golgi, the question arose, for each of these lineages as to whether the organelle (1) was ever present, (2) was present but is no longer a feature of the cellular configuration, or (3) is present but has been. Phylogenetic analysis to determine the evolutionary relationships of these organisms has placed them as embedded within various different eukaryotic groups, in almost every case having relatives with canonical stacked Golgi, rather than related to other organisms lacking stacks. Furthermore, in every case yet examined, when genome-scale data became available, genes were identified that encode orthologues of proteins that function at the golgi in mammalian and yeast systems. Localization data and functional assays have also confirmed that these proteins are expressed and indeed have shown that discrete golgi, of morphologies other than stacked cisternae, exist in several of these lineages. Recent genomic data for diverse eukaryotes, including from additional organisms with evidence for unstacked Golgi, therefore present the opportunity to understanding the evolution of Golgi structure across the broadest span of eukaryotes and organelle morphologies.
Keywords, golgi cell biologysecretionmembrane trafficking, background. At the intersection of the secretory and endocytic membrane-trafficking pathways in eukaryotes lies the golgi. This organelle comprises a series of compartments termed cisternae, providing a platform for protein transport, glycosylation, and targeting. The golgi is crucially important for normal cellular function, as demonstrated by the myriad diseases that result when genes associated with it are mutated. The most salient hallmark of Golgi structure is the presence of multiple membranous compartments, differentiated into cis, medial, and trans -golgi, and organized into flattened stacks, which quality facilitates many key golgi functions in mammalian cells. In mammalian cells, numerous proteins are involved in maintaining the structure and positioning of the golgi, as well as the specificity of membrane trafficking pathways to and from the golgi 3, although the precise mechanism of Golgi stacking is unknown. Golgins and Golgi reassembly and stacking proteins (grasps) are the main factors implicated in Golgi organization and stacking, as reviewed previously. The golgins are a collection of 11 proteins in mammalian cells defined by the presence of coiled-coil domains, attachment to golgi membranes near their C-termini (either by tail-anchor transmembrane domains or through binding to small gtpases and functions that include tethering/scaffolding 3,. The domain topology and functions of mammalian golgins have been reviewed extensively elsewhere 3,. Striking evidence for a role of grasp55, grasp65, gm130, and golgin-45 in stacking was shown by a knock-sideways experiment demonstrating that ectopic expression of grasp55 on mitochondria is sufficient for the stacking of mitochondrial and Golgi membranes together. A similar ectopic expression of golgin-84 on mitochondrial membranes also caused stacking of mitochondria. In addition to apparent roles in stacking, golgins, including GM130 and golgin-84, are involved in tethering specific transport vesicles destined for different regions of the golgi.
Camillo, golgi - wikipedia
Received:, accepted:, abstract, background, the golgi apparatus is a central meeting point for the endocytic and exocytic systems in eukaryotic cells, and the organelles dysfunction results in human disease. Its characteristic morphology of multiple online differentiated compartments organized into stacked flattened cisternae is one of the most recognizable features of modern eukaryotic cells, and yet how this is maintained is not well understood. The golgi is also an ancient aspect of eukaryotes, but the extent and nature of its complexity in the ancestor of eukaryotes is unclear. Various proteins have roles in organizing the golgi, chief among them being the golgins. Results, we address Golgi evolution by analyzing genome sequences from organisms which have lost stacked cisternae as a feature of their Golgi and those that have not. Using genomics and immunomicroscopy, we first identify golgi in the anaerobic amoeba. We then searched 87 genomes spanning eukaryotic diversity for presence of the most prominent proteins implicated in Golgi structure, focusing on golgins. We show some candidates as animal specific and others as ancestral to eukaryotes. Conclusions, none of the proteins examined show a phyletic distribution that correlates with the morphology of stacked cisternae, suggesting the possibility of stacking as an emergent property. Strikingly, however, the combination of golgins conserved among diverse eukaryotes allows for the most detailed reconstruction of the organelle to date, showing a sophisticated Golgi with differentiated compartments and trafficking pathways in the common eukaryotic ancestor.