Proteomic identification of heterogeneous nuclear ribonucleoprotein L as a novel component of SLM/Sam68 nuclear bodies

Background: Active pre-mRNA splicing occurs co-transcriptionally, and takes place throughout the nucleoplasm of eukaryotic cells. Splicing decisions are controlled by networks of nuclear RNA-binding proteins and their target sequences, sometimes in response to signalling pathways. Sam68 (Src-associated in mitosis 68 kDa) is the prototypic member of the STAR (Signal Transduction and Activation of RNA) family of RNA-binding proteins, which regulate splicing in response to signalling cascades. Nuclear Sam68 protein is concentrated within subnuclear organelles called SLM/Sam68 Nuclear Bodies (SNBs), which also contain some other splicing regulators, signalling components and nucleic acids. Results: We used proteomics to search for the major interacting protein partners of nuclear Sam68. In addition to Sam68 itself and known Sam68-associated proteins (heterogeneous nuclear ribonucleoproteins hnRNP A1, A2/B1 and G), we identified hnRNP L as a novel Sam68-interacting protein partner. hnRNP L protein was predominantly present within small nuclear protein complexes approximating to the expected size of monomers and dimers, and was quantitatively associated with nucleic acids. hnRNP L spatially co-localised with Sam68 as a novel component of SNBs and was also observed within the general nucleoplasm. Localisation within SNBs was highly specific to hnRNP L and was not shared by the closely-related hnRNP LL protein, nor any of the other Sam68-interacting proteins we identified by proteomics. The interaction between Sam68 and hnRNP L proteins was observed in a cell line which exhibits low frequency of SNBs suggesting that this association also takes place outside SNBs. Although ectopic expression of hnRNP L and Sam68 proteins independently affected splicing of CD44 variable exon v5 and TJP1 exon 20 minigenes, these proteins did not, however, co-operate with each other in splicing regulation of these target exons. Conclusion: Here we identify hnRNP L as a novel SNB component. We show that, compared with other identified Sam68-associated hnRNP proteins and hnRNP LL, this co-localisation within SNBs is specific to hnRNP L. Our data suggest that the novel Sam68-hnRNP L protein interaction may have a distinct role within SNBs

Challenges in estimating soil water

[Introduction]: Most of Australia’s dryland cropping is characterised by unreliable rainfall with frequent long gaps between falls. Stored soil water is therefore essential to support crop growth during the growing season while water stored during fallows has varying importance, depending on soil type and rainfall patterns in relation to cropping periods. For example, a winter crop at Walpeup in Victoria derives 10% of its water supply from soil water at planting while a winter crop at Emerald will access 80% of its water supply from stored soil water (Thomas et al 2007). Even when dependence on stored water is small, extra water can make a valuable difference to crop yield and profitability, especially in typical dry-finish seasons (Kirkegaard et al 2014). An understanding of available water before a crop is planted can influence management decisions (area planted, fertilizer rates). Estimating plant available water (PAW) also requires an estimate of a soils ability to store water, its plant available water capacity (PAWC). This paper presents some observations of soil water from a 17-year study comparing water balances (runoff, evaporation and deep drainage) for a set of small contour bay catchments near Roma in southern Queensland. Our aim is to demonstrate some of the challenges associated with field measurement of both PAWC and PAW. This analysis is an extension of a detailed description of the development of SoilWaterApp (Freebairn et al. 2018)

Combining biophysical and price simulations to assess the economics of long-term crop rotations

Long-run rotational gross margins were calculated with yields derived from biophysical simulations in APSIM over a period of 100+ years and prices simulated in @Risk based on subjective triangular price distributions elicited from the Jimbour Plains farmer group. Rotations included chickpeas, cotton, lucerne, sorghum, wheat and different lengths of fallow. Output presented to the farmers included mean annual GMs and distributions of GMs with box and whisker plots found to be suitable. Mean-standard deviation and first and second-degree stochastic dominance efficiency measures were also calculated. Including lucerne in the rotations improved some sustainability indicators but reduced profitability.Crop Production/Industries, Farm Management,

A study of preferences from nineteen selections in current children's literature.

Thesis (Ed.M.)--Boston Universit

Striving for autonomy : representative female characters in the detective novels of P. D. James : a thesis presented in partial fulfilment of the requirements for the degree of Master of Arts in English at Massey University, Palmerston North, New Zealand

Representative female characters from several of P D James's detective novels are used to exemplify the changes in women's position in society during the four decades (from the early 1960s to the late 1990s) which span James's publishing career and which coincide with the period known as the second wave of feminism. Women characters have always taken a prominent place in P D James's detective fiction, and since the 1970s her books have increasingly foregrounded the problems that women have when working in male-dominated professions, revealing their increasing autonomy but also disclosing the continuing limitations of that autonomy. Her novels are acknowledged as becoming increasingly literary. In her early novels James followed the formula of the classic detective fiction genre quite closely. During the 1970s she experimented with novels that on the surface read as detective novels, while functioning subtextually in relation to myths and metaphors. In her most recent works she transcends the genre, using the detective formula simply as a framework for her novels of literary realism

A novel androgen-regulated isoform of the TSC2 tumour suppressor gene increases cell proliferation

TSC2 (Tuberous sclerosis complex 2) is an important tumour suppressor gene, mutations within which are linked to the development of tuberous sclerosis and implicated in multiple tumour types. TSC2 protein complexes with TSC1 and blocks the ability of the Rheb (Ras homolog enriched in brain) GTPase to activate mTOR (mammalian target of rapamycin), a crucial signal transducer which regulates protein synthesis and cell growth. Here, we report the characterisation of a novel isoform of TSC2 which is under direct control of the ligand-activated androgen receptor. TSC2 isoform A (TSC2A) is derived from an internal androgen-regulated alternative promoter and encodes a 508-amino acid cytoplasmic protein corresponding to the C-terminal region of full-length TSC2, lacking the interaction domain for TSC1 and containing an incomplete interaction domain required for Rheb inactivation. Expression of TSC2A is induced in response to androgens and full-length TSC2 is co-ordinately down-regulated, indicating an androgen-driven switch in TSC2 protein isoforms. In contrast to the well-characterised suppressive effect on cell proliferation of full-length TSC2 protein, both LNCaP and HEK293 cells over-expressing TSC2 isoform A proliferate more rapidly (measured by MTT assays) and have increased levels of cells in S-phase (measured by both Edu staining and FACS analysis). Our work indicates, for the first time, a novel role for this well-known tumour suppressor gene, which encodes an activator of cell proliferation in response to androgen stimulation

Recording and reporting practices in some New England kindergartens.

Thesis (M.A.)--Boston Universit

On the mechanical properties of N-functionalised dipeptide gels

The properties of a hydrogel are controlled by the underlying network that immobilizes the solvent. For gels formed by the self-assembly of a small molecule, it is common to show the primary fibres that entangle to form the network by microscopy, but it is difficult to access information about the network. One approach to understand the network is to examine the effect of the concentration on the rheological properties, such that G cx, where G is the storage modulus and c is the concentration. A number of reports link the exponent x to a specific type of network. Here, we discuss a small library of gels formed using functionalized dipeptides, and describe the underlying networks of these gels, using microscopy, small angle scattering and rheology. We show that apparently different networks can give very similar values of x