Wnt evolution and function shuffling in liberal and conservative chordate genomes

Background What impact gene loss has on the evolution of developmental processes, and how function shuffling has affected retained genes driving essential biological processes, remain open questions in the fields of genome evolution and EvoDevo. To investigate these problems, we have analyzed the evolution of the Wnt ligand repertoire in the chordate phylum as a case study. Results We conduct an exhaustive survey of Wnt genes in genomic databases, identifying 156 Wnt genes in 13 non-vertebrate chordates. This represents the most complete Wnt gene catalog of the chordate subphyla and has allowed us to resolve previous ambiguities about the orthology of many Wnt genes, including the identification of WntA for the first time in chordates. Moreover, we create the first complete expression atlas for the Wnt family during amphioxus development, providing a useful resource to investigate the evolution of Wnt expression throughout the radiation of chordates. Conclusions Our data underscore extraordinary genomic stasis in cephalochordates, which contrasts with the liberal and dynamic evolutionary patterns of gene loss and duplication in urochordate genomes. Our analysis has allowed us to infer ancestral Wnt functions shared among all chordates, several cases of function shuffling among Wnt paralogs, as well as unique expression domains for Wnt genes that likely reflect functional innovations in each chordate lineage. Finally, we propose a potential relationship between the evolution of WntA and the evolution of the mouth in chordates

An inter-subunit protein-peptide interface that stabilizes the specific activity and oligomerization of the AAA+ chaperone Reptin

The work was supported by: the Czech Science Foundation 16-20860S (PM, LH) and 16-07321S (BV, TH), the project MEYS – NPS I – LO1413, and MH CZ - DRO (MMCI, 00209805); the BBSRC RASOR consortium (BB/C511599/1; United Kingdom); Cancer Research UK (C21383/A6950); The International Centre for Cancer Vaccine Science project carried out within the International Research Agendas programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund; A*STAR, Singapore and NSCC, Singapore.Reptin is a member of the AAA+ superfamily whose members can exist in equilibrium between monomeric apo forms and ligand bound hexamers. Inter-subunit protein-protein interfaces that stabilize Reptin in its oligomeric state are not well-defined. A self-peptide binding assay identified a protein-peptide interface mapping to an inter-subunit “rim” of the hexamer bridged by Tyrosine-340. A Y340A mutation reduced ADP-dependent oligomer formation using a gel filtration assay, suggesting that Y340 forms a dominant oligomer stabilizing side chain. The monomeric ReptinY340A mutant protein exhibited increased activity to its partner protein AGR2 in an ELISA assay, further suggesting that hexamer formation can preclude certain protein interactions. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) demonstrated that the Y340A mutation attenuated deuterium suppression of Reptin in this motif in the presence of ligand. By contrast, the tyrosine motif of Reptin interacts with a shallower pocket in the hetero-oligomeric structure containing Pontin and HDX-MS revealed no obvious role of the Y340 side chain in stabilizing the Reptin-Pontin oligomer. Molecular dynamic simulations (MDS) rationalized how the Y340A mutation impacts upon a normally stabilizing inter-subunit amino acid contact. MDS also revealed how the D299N mutation can, by contrast, remove oligomer de-stabilizing contacts. These data suggest that the Reptin interactome can be regulated by a ligand dependent equilibrium between monomeric and hexameric forms through a hydrophobic inter-subunit protein-protein interaction motif bridged by Tyrosine-340. Significance Discovering dynamic protein-protein interactions is a fundamental aim of research in the life sciences. An emerging view of protein-protein interactions in higher eukaryotes is that they are driven by small linear polypeptide sequences; the linear motif. We report on the use of linear-peptide motif screens to discover a relatively high affinity peptide-protein interaction for the AAA+ and pro-oncogenic protein Reptin. This peptide interaction site was shown to form a ‘hot-spot’ protein-protein interaction site, and validated to be important for ligand-induced oligomerization of the Reptin protein. These biochemical data provide a foundation to understand how single point mutations in Reptin can impact on its oligomerization and protein-protein interaction landscape.PostprintPeer reviewe

Mapping the H2 resistance effective against Globodera pallida pathotype Pa1 in tetraploid potato

This work was supported by the Rural & Environment Science & Analytical Services Division of the Scottish Government, the BBSRC, through the joint projects CRF/2009/SCRI/SOP 0929, BB/L008025/1 and BB/K018299/1. Additional funding was obtained through the James Hutton Institute SEEDCORN initiative, AHDB Potato, the Perry Foundation and The Felix Cobbold Trust. Amanpreet Kaur was supported by the Commonwealth Scholarship Commission through a Commonwealth split-site Ph.D. grant.Key message: The nematode resistance gene H2 was mapped to the distal end of chromosome 5 in tetraploid potato. The H2 resistance gene, introduced into cultivated potatoes from the wild diploid species Solanum multidissectum, confers a high level of resistance to the Pa1 pathotype of the potato cyst nematode Globodera pallida. A cross between tetraploid H2-containing breeding clone P55/7 and susceptible potato variety Picasso yielded an F1 population that segregated approximately 1:1 for the resistance phenotype, which is consistent with a single dominant gene in a simplex configuration. Using genome reduction methodologies RenSeq and GenSeq, the segregating F1 population enabled the genetic characterisation of the resistance through a bulked segregant analysis. A diagnostic RenSeq analysis of the parents confirmed that the resistance in P55/7 cannot be explained by previously characterised resistance genes. Only the variety Picasso contained functionally characterised disease resistance genes Rpi-R1, Rpi-R3a, Rpi-R3b variant, Gpa2 and Rx, which was independently confirmed through effector vacuum infiltration assays. RenSeq and GenSeq independently identified sequence polymorphisms linked to the H2 resistance on the top end of potato chromosome 5. Allele-specific KASP markers further defined the locus containing the H2 gene to a 4.7 Mb interval on the distal short arm of potato chromosome 5 and to positions that correspond to 1.4 MB and 6.1 MB in the potato reference genome.Publisher PDFPeer reviewe

Distance measurement of a noncovalently bound Y@C82 pair with double electron electron resonance spectroscopy

Paramagnetic endohedral fullerenes with long spin coherence times, such as N@C60 and Y@C82, are being explored as potential spin quantum bits (qubits). Their use for quantum information processing requires a way to hold them in fixed spatial arrangements. Here we report the synthesis of a porphyrinbased two-site receptor 1, offering a rigid structure that binds spin-active fullerenes (Y@C82) at a center-to-center distance of 5.0 nm, predicted from molecular simulations. The spin-spin dipolar coupling was measured with the pulsed EPR spectroscopy technique of double electron electron resonance (DEER) and analysed to give a distance of 4.87 nm with a small distribution of distances

Voice Therapy For Transgender People

Transrodne osobe su osobe koje osjećaju nesklad između roda s kojim se poistovjećuju i spola u kojem su se rodili. Transrodnost je širok pojam i obuhvaća čitav spektar rodno nenormativnih identiteta. Dva glavna obrasca tranzicije prema drugom spolu, odnosno rodu su: Male-to-Female (MtF) i Female-to-Male (FtM). Transrodne osobe navode da ih njihov glas često “izdaje”, odnosno otkriva njihov biološki rod te predstavlja posljednju prepreku u potpuno uživljavanje u novu rodnu ulogu. Bez obzira na to, često nisu svjesni da se zbog glasa mogu obratiti logopedu. Cilj ovog preglednog rada je dati uvid u poremećaje glasa kod transrodnih osoba, odnosno opisati logopedsku procjenu te terapijske postupke, a sve u svrhu boljeg razumijevanja potreba ovih osoba.Transgender individuals are people who feel an incongruity between their self-identified gender and their birth gender. Transgenderism is a broad term and includes a spectrum of gender-nonconforming identities. The two main patterns of gender transition are Male-to-Female (MtF) and Female-to-Male (FtM).Transgender people often think their voice “betrays” them, i.e. it reveals their biological gender and represents the last obstacle to the individual’s full enjoyment of his/her new gender role. Regardless of this, many transgender individuals are not aware of the fact that they can go to speech and language pathologists (SLP) for therapy. The aim of this review article is to provide insight into the voice disorders of transgender people, i.e. to describe the assessment process and therapy protocols in hope of gaining a better understanding of the needs of this population