School of BioSciences - Research Publications

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    C19ORF84 connects piRNA and DNA methylation machineries to defend the mammalian germ line.
    Zoch, A ; Konieczny, G ; Auchynnikava, T ; Stallmeyer, B ; Rotte, N ; Heep, M ; Berrens, RV ; Schito, M ; Kabayama, Y ; Schöpp, T ; Kliesch, S ; Houston, B ; Nagirnaja, L ; O'Bryan, MK ; Aston, KI ; Conrad, DF ; Rappsilber, J ; Allshire, RC ; Cook, AG ; Tüttelmann, F ; O'Carroll, D (Elsevier BV, 2024-03-21)
    In the male mouse germ line, PIWI-interacting RNAs (piRNAs), bound by the PIWI protein MIWI2 (PIWIL4), guide DNA methylation of young active transposons through SPOCD1. However, the underlying mechanisms of SPOCD1-mediated piRNA-directed transposon methylation and whether this pathway functions to protect the human germ line remain unknown. We identified loss-of-function variants in human SPOCD1 that cause defective transposon silencing and male infertility. Through the analysis of these pathogenic alleles, we discovered that the uncharacterized protein C19ORF84 interacts with SPOCD1. DNMT3C, the DNA methyltransferase responsible for transposon methylation, associates with SPOCD1 and C19ORF84 in fetal gonocytes. Furthermore, C19ORF84 is essential for piRNA-directed DNA methylation and male mouse fertility. Finally, C19ORF84 mediates the in vivo association of SPOCD1 with the de novo methylation machinery. In summary, we have discovered a conserved role for the human piRNA pathway in transposon silencing and C19ORF84, an uncharacterized protein essential for orchestrating piRNA-directed DNA methylation.
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    Acetylcholine esterase of Drosophila melanogaster: a laboratory model to explore insecticide susceptibility gene drives
    Hernandes, N ; Qi, XM ; Bhide, S ; Brown, C ; Camm, BJ ; Baxter, SW ; Robin, C (JOHN WILEY & SONS LTD, 2024-06)
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    Defining and Reducing Variant Classification Disparities.
    Dawood, M ; Fayer, S ; Pendyala, S ; Post, M ; Kalra, D ; Patterson, K ; Venner, E ; Muffley, LA ; Fowler, DM ; Rubin, AF ; Posey, JE ; Plon, SE ; Lupski, JR ; Gibbs, RA ; Starita, LM ; Robles-Espinoza, CD ; Coyote-Maestas, W ; Gallego Romero, I (Cold Spring Harbor Laboratory, 2024-04-12)
    BACKGROUND: Multiplexed Assays of Variant Effects (MAVEs) can test all possible single variants in a gene of interest. The resulting saturation-style data may help resolve variant classification disparities between populations, especially for variants of uncertain significance (VUS). METHODS: We analyzed clinical significance classifications in 213,663 individuals of European-like genetic ancestry versus 206,975 individuals of non-European-like genetic ancestry from All of Us and the Genome Aggregation Database. Then, we incorporated clinically calibrated MAVE data into the Clinical Genome Resource's Variant Curation Expert Panel rules to automate VUS reclassification for BRCA1, TP53, and PTEN . RESULTS: Using two orthogonal statistical approaches, we show a higher prevalence ( p ≤5.95e-06) of VUS in individuals of non-European-like genetic ancestry across all medical specialties assessed in all three databases. Further, in the non-European-like genetic ancestry group, higher rates of Benign or Likely Benign and variants with no clinical designation ( p ≤2.5e-05) were found across many medical specialties, whereas Pathogenic or Likely Pathogenic assignments were higher in individuals of European-like genetic ancestry ( p ≤2.5e-05). Using MAVE data, we reclassified VUS in individuals of non-European-like genetic ancestry at a significantly higher rate in comparison to reclassified VUS from European-like genetic ancestry ( p =9.1e-03) effectively compensating for the VUS disparity. Further, essential code analysis showed equitable impact of MAVE evidence codes but inequitable impact of allele frequency ( p =7.47e-06) and computational predictor ( p =6.92e-05) evidence codes for individuals of non-European-like genetic ancestry. CONCLUSIONS: Generation of saturation-style MAVE data should be a priority to reduce VUS disparities and produce equitable training data for future computational predictors.
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    Genetic mutation of Cep76 results in male infertility due to abnormal sperm tail composition
    Houston, BJ ; Merriner, DJ ; Stathatos, GG ; Nguyen, JH ; O'Connor, AE ; Lopes, AM ; Conrad, DF ; Baker, M ; Dunleavy, JEM ; O'Bryan, MK (Life Science Alliance LLC, 2024-06)
    The transition zone is a specialised gate at the base of cilia/flagella, which separates the ciliary compartment from the cytoplasm and strictly regulates protein entry. We identified a potential new regulator of the male germ cell transition zone, CEP76. We demonstrated that CEP76 was involved in the selective entry and incorporation of key proteins required for sperm function and fertility into the ciliary compartment and ultimately the sperm tail. In the mutant, sperm tails were shorter and immotile as a consequence of deficits in essential sperm motility proteins including DNAH2 and AKAP4, which accumulated at the sperm neck in the mutant. Severe annulus, fibrous sheath, and outer dense fibre abnormalities were also detected in sperm lacking CEP76. Finally, we identified that CEP76 dictates annulus positioning and structure. This study suggests CEP76 as a male germ cell transition zone protein and adds further evidence to the hypothesis that the spermatid transition zone and annulus are part of the same functional structure.
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    Gut microbiota in the short-beaked echidna (Tachyglossus Aculeatus) shows stability across gestation
    Buthgamuwa, I ; Fenelon, JC ; Roser, A ; Meer, H ; Johnston, SD ; Dungan, AM (Wiley, 2023-12)
    Indigenous gut microbial communities (microbiota) play critical roles in health and may be especially important for the mother and fetus during pregnancy. Monotremes, such as the short-beaked echidna, have evolved to lay and incubate an egg, which hatches in their pouch where the young feeds. Since both feces and eggs pass through the cloaca, the fecal microbiota of female echidnas provides an opportunity for vertical transmission of microbes to their offspring. Here, we characterize the gut/fecal microbiome of female short-beaked echidnas and gain a better understanding of the changes that may occur in their microbiome as they go through pregnancy. Fecal samples from four female and five male echidnas were obtained from the Currumbin Wildlife Sanctuary in Queensland and sequenced to evaluate bacterial community structure. We identified 25 core bacteria, most of which were present in male and female samples. Genera such as Fusobacterium, Bacteroides, Escherichia-Shigella, and Lactobacillus were consistently abundant, regardless of sex or gestation stage, accounting for 58.00% and 56.14% of reads in male and female samples, respectively. The echidna microbiome remained stable across the different gestation stages, though there was a significant difference in microbiota composition between male and female echidnas. This study is the first to describe the microbiome composition of short-beaked echidnas across reproductive phases and allows the opportunity for this novel information to be used as a metric of health to aid in the detection of diseases triggered by microbiota dysbiosis.
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    Ecomorphological correlates of inner ear shape in Australian limb-reduced skinks (Scincidae: Sphenomorphini)
    Camaiti, M ; Wiles, J ; Aguilar, R ; Hutchinson, MN ; Hipsley, CA ; Chapple, DG ; Evans, AR (OXFORD UNIV PRESS, 2023-12-01)
    Abstract The inner ear labyrinth is an organ able to perceive balance and spatial orientation, but the drivers of its morphological variation across and within vertebrate lineages are unclear. We assess two competing hypotheses whether this organ, and specifically the semicircular canals, modifies its shape as a functional adaptation to ecology and locomotion, or according to the constraints of skull morphology. We test these using 52 species of Australian sphenomorphines, a group of scincid lizards that evolved changes in body shape and locomotory adaptations to fossoriality multiple times independently, by reducing their limbs. We find a correlation between semicircular canal shape and degree of limb reduction in these lizards, supporting a functional hypothesis. The interaction between body shape and substrate ecology is also a significant predictor. The wider and more eccentric semicircular canals of limb-reduced skinks indicate higher balance sensitivity and manoeuvrability compared with fully limbed skinks, probably as an adaptation to navigating cluttered environments. Conversely, our results show only a minimal influence of skull constraints on semicircular canal shape, having instead significant effects on size. This supports the hypothesis that in these skinks inner ear shape evolution is driven by specific locomotory strategies more than it is constrained by cranial anatomy.
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    A targeted approach to enrich host-associated bacteria for metagenomic sequencing.
    Dungan, AM ; Tandon, K ; Jameson, V ; Gotze, CR ; Blackall, LL ; van Oppen, MJH (Oxford University Press (OUP), 2024)
    Multicellular eukaryotic organisms are hosts to communities of bacteria that reside on or inside their tissues. Often the eukaryotic members of the system contribute to high proportions of metagenomic sequencing reads, making it challenging to achieve sufficient sequencing depth to evaluate bacterial ecology. Stony corals are one such complex community; however, separation of bacterial from eukaryotic (primarily coral and algal symbiont) cells has so far not been successful. Using a combination of hybridization chain reaction fluorescence in situ hybridization and fluorescence activated cell sorting (HCR-FISH + FACS), we sorted two populations of bacteria from five genotypes of the coral Acropora loripes, targeting (i) Endozoicomonas spp, and (ii) all other bacteria. NovaSeq sequencing resulted in 67-91 M reads per sample, 55%-90% of which were identified as bacterial. Most reads were taxonomically assigned to the key coral-associated family, Endozoicomonadaceae, with Vibrionaceae also abundant. Endozoicomonadaceae were 5x more abundant in the 'Endozoicomonas' population, highlighting the success of the dual-labelling approach. This method effectively enriched coral samples for bacteria with <1% contamination from host and algal symbionts. The application of this method will allow researchers to decipher the functional potential of coral-associated bacteria. This method can also be adapted to accommodate other host-associated communities.
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    Tissue-associated and vertically transmitted bacterial symbiont in the coral Pocillopora acuta
    Maire, J ; Tsang Min Ching, SJ ; Damjanovic, K ; Epstein, HE ; Judd, LM ; Blackall, LL ; van Oppen, MJH (OXFORD UNIV PRESS, 2024-01-08)
    Coral microhabitats are colonized by a myriad of microorganisms, including diverse bacteria which are essential for host functioning and survival. However, the location, transmission, and functions of individual bacterial species living inside the coral tissues remain poorly studied. Here, we show that a previously undescribed bacterial symbiont of the coral Pocillopora acuta forms cell-associated microbial aggregates (CAMAs) within the mesenterial filaments. CAMAs were found in both adults and larval offspring, suggesting vertical transmission. In situ laser capture microdissection of CAMAs followed by 16S rRNA gene amplicon sequencing and shotgun metagenomics produced a near complete metagenome-assembled genome. We subsequently cultured the CAMA bacteria from Pocillopora acuta colonies, and sequenced and assembled their genomes. Phylogenetic analyses showed that the CAMA bacteria belong to an undescribed Endozoicomonadaceae genus and species, which we propose to name Candidatus Sororendozoicomonas aggregata gen. nov sp. nov. Metabolic pathway reconstruction from its genome sequence suggests this species can synthesize most amino acids, several B vitamins, and antioxidants, and participate in carbon cycling and prey digestion, which may be beneficial to its coral hosts. This study provides detailed insights into a new member of the widespread Endozoicomonadaceae family, thereby improving our understanding of coral holobiont functioning. Vertically transmitted, tissue-associated bacteria, such as Sororendozoicomonas aggregata may be key candidates for the development of microbiome manipulation approaches with long-term positive effects on the coral host.
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    Plastid phylogenomics of the Eriostemon group (Rutaceae; Zanthoxyloideae): support for major clades and investigation of a backbone polytomy
    Orel, H ; McLay, TGB ; Neal, WC ; Forster, PI ; Bayly, MJ ; Bruhl, J (CSIRO Publishing, 2023-08-22)
    Most of Australia’s sclerophyllous Rutaceae belong to a clade informally known as the ‘Eriostemon group’ (including 16 genera, ~209 species). We investigated generic relationships in this group using analyses of complete plastome sequence data for 60 species and analyses of a supermatrix including sequences of four plastome spacer regions for 22 additional species. Maximum likelihood, Bayesian inference, and shortcut coalescent phylogenetic analyses produced congruent phylogenies that were highly supported, except for a series of short unsupported branches in the backbone of the Eriostemon group. We found high support for four major clades branching from this polytomy and discuss evolutionary inferences of generic relationships in each lineage. In an effort to resolve the polytomy, we analysed gene tree topologies in tree space, phylogenetic informativeness with likelihood mapping, and conducted topology tests to assess support for all possible topological resolutions of the polytomy. These approaches did not clarify the polytomy, which may be caused by insufficient data, features of plastome evolution, or rapid radiation. Results from analyses of the combined supermatrix dataset suggest that Philotheca section Philotheca is paraphyletic with regards to Drummondita and Geleznowia. In all phylogenies, Philotheca sections Corynonema and Cyanochlamys were not placed with other members of Philotheca.
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    Discovery of novel neutral glycosphingolipids in cereal crops: rapid profiling using reversed-phased HPLC-ESI-QqTOF with parallel reaction monitoring
    Yu, D ; Boughton, BA ; Rupasinghe, TWT ; Hill, CB ; Herrfurth, C ; Scholz, P ; Feussner, I ; Roessner, U (Nature Portfolio, 2023-12-19)
    This study explores the sphingolipid class of oligohexosylceramides (OHCs), a rarely studied group, in barley (Hordeum vulgare L.) through a new lipidomics approach. Profiling identified 45 OHCs in barley (Hordeum vulgare L.), elucidating their fatty acid (FA), long-chain base (LCB) and sugar residue compositions; and was accomplished by monophasic extraction followed by reverse-phased high performance liquid chromatography electrospray ionisation quadrupole-time-of-flight tandem mass spectrometry (HPLC-ESI-QqTOF-MS/MS) employing parallel reaction monitoring (PRM). Results revealed unknown ceramide species and highlighted distinctive FA and LCB compositions when compared to other sphingolipid classes. Structurally, the OHCs featured predominantly trihydroxy LCBs associated with hydroxylated FAs and oligohexosyl residues consisting of two-five glucose units in a linear 1 → 4 linkage. A survey found OHCs in tissues of major cereal crops while noting their absence in conventional dicot model plants. This study found salinity stress had only minor effects on the OHC profile in barley roots, leaving questions about their precise functions in plant biology unanswered.