Identification of a Sam68 Ribonucleoprotein Complex Regulated by Epidermal Growth Factor

Sam68, Src associated in mitosis of 68 kDa, is a known RNA-binding protein and a signaling adaptor protein for tyrosine kinases. However, the proteins associated with Sam68 and the existence of a Sam68 complex, its mass, and regulation are, however, unknown. Herein we identify a large Sam68 complex with a mass >1 MDa in HeLa cells that is composed of ∼40 proteins using an immunoprecipitation followed by a mass spectrometry approach. Many of the proteins identified are RNA-binding proteins and are known components of a previously identified structure termed the spreading initiation center. The large Sam68 complex is a ribonucleoprotein complex, as treatment with RNases caused a shift in the molecular mass of the complex to 200–450 kDa. Moreover, treatment of HeLa cells with phorbol 12-myristate 13-acetate or epidermal growth factor induced the disassociation of Sam68 from the large complex and the appearance of Sam68 within the smaller complex. Actually, in certain cell lines such as breast cancer cell lines MCF-7 and BT-20, Sam68 exists in equilibrium between a large and a small complex. The appearance of the small Sam68 complex in cells correlates with the ability of Sam68 to promote the alternative splicing of CD44 and cell migration. Our findings show that Sam68 exists in equilibrium in transformed cells between two complexes and that extracellular signals, such as epidermal growth factor stimulation, promote alternative splicing by modulating the composition of the Sam68 complex.     

Blood levels of kynurenines, interleukin-23 and soluble human leucocyte antigen-G at different stages of Huntington's disease

There is substantial evidence that abnormal concentrations of oxidised tryptophan metabolites, produced via the kynurenine pathway, contribute to progressive neurodegeneration in Huntington's disease. We have now examined the blood levels of these metabolites in patients at different stages of Huntington's disease, assessed both in terms of clinical disease severity and numbers of CAG repeats. Close relatives of the patients were included in the study as well as unrelated healthy controls. Levels of lipid peroxidation products, the pro-inflammatory cytokine interleukin (IL)-23 and the soluble human leucocyte antigen-G (sHLA-G) were also measured. There were lower levels of tryptophan and a higher kynurenine : tryptophan ratio, indicating activation of indoleamine-2,3-dioxygenase, in the most severely affected group of patients, with increased levels of IL-23 and sHLA-G. Marked correlations were noted between IL-23 and the patient severity group, anthranilic acid levels and the number of CAG repeats, and between anthranilic acid and IL-23, supporting our previous evidence of a relationship between anthranilic acid and inflammatory status. Tryptophan was negatively correlated with symptom severity and number of CAG repeats, and positively correlated with sHLA-G. The results support the proposal that tryptophan metabolism along the kynurenine pathway in Huntington's disease is related to the degree of genetic abnormality, to clinical disease severity and to aspects of immunopathogenesis.     

Competition and Cooperation among Relational Memory Representations

Mnemonic processing engages multiple systems that cooperate and compete to support task performance. Exploring these systems’ interaction requires memory tasks that produce rich data with multiple patterns of performance sensitive to different processing sub-components. Here we present a novel context-dependent relational memory paradigm designed to engage multiple learning and memory systems. In this task, participants learned unique face-room associations in two distinct contexts (i.e., different colored buildings). Faces occupied rooms as determined by an implicit gender-by-side rule structure (e.g., male faces on the left and female faces on the right) and all faces were seen in both contexts. In two experiments, we use behavioral and eye-tracking measures to investigate interactions among different memory representations in both younger and older adult populations; furthermore we link these representations to volumetric variations in hippocampus and ventromedial PFC among older adults. Overall, performance was very accurate. Successful face placement into a studied room systematically varied with hippocampal volume. Selecting the studied room in the wrong context was the most typical error. The proportion of these errors to correct responses positively correlated with ventromedial prefrontal volume. This novel task provides a powerful tool for investigating both the unique and interacting contributions of these systems in support of relational memory.     

Haemoglobin Belfast 15 (A12) Tryptophan→Arginine: A New Unstable Haemoglobin Variant

A new unstable haemoglobin, α₂β₂15 Trp→ Arg (Hb Belfast), with increased oxygen affinity has been found during the routine investigation of a long-stay psychiatric patient. It seemed to cause little haematological disorder. The reticulocytes synthesized normal and abnormal β-chains at the same rate but in the circulating blood Hb Belfast amounted to only 27·5% of the total haemoglobin.     

Biosynthetic pathway of desmosines in elastin

1. Elastins purified by various methods from the ligamentum nuchae and the lungs of cattle of various ages were analysed for amino acid compositions and lysine-derived cross-links. 2. In fully mature elastins from adults the main cross-links were desmosine, isodesmosine and lysinonorleucine and the so-called aldol-condensation product. Trace amounts of merodesmosine were also found. 3. During the normal maturation of elastins the amounts of desmosine, isodesmosine and lysinonorleucine increased, whereas the aldol-condensation product and intact lysine residues decreased and merodesmosine remained the same. 4. Elastins from young animals contained significant amounts of dehydromerodesmosine whereas elastins from adults contained virtually nil. Evidence is presented which suggests that the biosynthetic pathway of desmosine and isodesmosine proceeds via the aldol-condensation product and dehydromerodesmosine.     

Palmityl-CoA oxidase: Detection in several guinea pig tissues and peroxisomal localisation in mucosa of small intestine

Using a sensitive assay palmityl-CoA oxidase activity was detected in several guinea pig tissues including liver, kidney, small intestine, lung, muscle, spleen and heart. Subcellular fractionation using sucrose and metrizamide gradients demonstrated that in small intestinal mucosa palmityl-CoA oxidase is localised in the same particles as catalase. These observations are discussed in the context of a role for peroxisomal fatty acid oxidation in energy balance and thermogenesis.