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Turner, S. C.

Publications and source records attributed to Turner, S. C..

3 recordsLinked to original sources

Establishing MS2-MCP-based single-molecule RNA visualization in Schizosaccharomyces pombe

Single-molecule RNA imaging using the MS2-MCP system has transformed the study of RNA biology across model organisms. However, this technology has remained unavailable for fission yeast (Schizosaccharomyces pombe), even though fission yeast is a central model for eukaryotic gene expression. Achieving single-molecule sensitivity requires identifying a narrow optimum where RNA labels are sufficiently bright while background fluorescence remains minimal. We have now accomplished this for S. pombe by systematically optimizing MCP expression and localization--screening a panel of constitutive S. pombe promoters and evaluating combinations of nuclear localization and export signals (NLSs and NESs). The resulting, successful constructs use tandem StayGold as the MCP fluorescent tag, taking advantage of its superior photostability. Together with optimized vectors for MS2 stem-loop tagging of endogenous transcripts, these tools enable single-molecule RNA imaging in fission yeast, opening the door to quantitative analyses of RNA dynamics in this core genetic model. Summary statementWe establish single-molecule RNA visualization with MS2/MCP for fission yeast, S. pombe, an important model organism to study RNA biology.

molecular biology↗

Plant neighbors differentially alter a focal species' biotic interactions through changes to resource allocation

Plant resource allocation strategies are thought to be largely a consequence of changing abiotic conditions and evolutionary history. However, biotic interactions also influence how a plant allocates resources. As a result, plants mediate indirect interactions between organisms above- and belowground through resource allocation. Neighboring plants can influence plant fitness directly through competition for resources, and indirectly by altering associated community interactions (associational effects). Given the importance of community interactions for plant success, and the known ability for plant neighbors to change these interactions, the goal of this "pandemic project" was to separate inter- and intraspecific plant associations, above- and belowground, to understand how different plant neighbors alter plant resource allocation, and if this in turn alters biotic interactions. We specifically investigated associational effects on herbivory and soil microbial community interactions. To do so, we established a common garden experiment, manipulating plant neighbors and extent of interactions (aboveground only versus above- and belowground interactions, using customized pot types), and measured changes to a focal plant and its biotic interactions over two growing seasons. We found evidence of both neighbor effects and pot type, showing that neighbor interactions affect a focal plant through both above- and belowground processes, and how the focal plant is affected depends on neighbor identity. Though neighbors did not directly alter herbivory or most soil microbial interactions, they did alter the relationship between belowground microbial communities and plant function. Resource allocation responses were reduced with time, showing the importance of extending experiments beyond a single growing season, and is an important consideration when making predictions about plant responses to changing conditions. This study contributes to a growing body of work showing how the community context affects the above- and belowground interactions of a plant through plant resource allocation strategies.

ecology↗

Passage of transmissible cancers in the Tasmanian devil is due to a dominant, shared peptide motif and a limited repertoire of MHC-I allotypes

Transmissible cancers are spread via the passage of malignant cells. The survival of the Tasmanian devil, the largest marsupial carnivore, is threatened by two independent transmissible cancers, devil facial tumour (DFT) 1 and devil facial tumour 2 (DFT2). To aid the development of a peptide vaccine and to interrogate how histocompatibility barriers can be overcome, we analysed the peptides bound to Major Histocompatibility Complex class I molecules from the Tasmanian devil and its transmissible tumours. Comparison of the peptidomes from DFT1+IFNγ, DFT2 and host fibroblast cells demonstrates a shared motif, despite differences in MHC-I allotypes between the cell lines. Importantly, DFT1+IFNγ and DFT2 share the presentation of peptides derived from neural proteins, reflecting a common cellular origin that should be exploited for vaccine design. These results suggest that some polymorphisms between tumours and host are ‘hidden’ by a common peptide motif, providing the potential for permissive passage of infectious cells.Competing Interest StatementThe authors have declared no competing interest.View Full Text

immunology↗