Spatial proteomics finds CD155 and Endophilin-A1 as mediators of growth and invasion in medulloblastoma

The composition of the plasma membrane (PM)-associated proteome of tumor cells determines cell-cell and cell-matrix interactions and the response to environmental cues. Whether the PM-associated proteome impacts the phenotype of Medulloblastoma (MB) tumor cells and how it adapts in response to growth factor cues is poorly understood. Using a spatial proteomics approach, we observed that hepatocyte growth factor (HGF)-induced activation of the receptor tyrosine kinase c-MET in MB cells changes the abundance of transmembrane and membrane-associated proteins. The depletion of MAP4K4, a pro-migratory effector kinase downstream of c-MET, leads to a specific decrease of the adhesion and immunomodulatory receptor CD155 and of components of the fast-endophilin-mediated endocytosis (FEME) machinery in the PM-associated proteome of HGF-activated MB cells. The decreased surface expression of CD155 or of the fast-endophilin-mediated endocytosis effector endophilin-A1 reduces growth and invasiveness of MB tumor cells in the tissue context. These data thus describe a novel function of MAP4K4 in the control of the PM-associated proteome of tumor cells and identified two downstream effector mechanisms controlling proliferation and invasiveness of MB cells

Proteini glutena – determinanta kvaliteta pšenice

Gluten is complex of gliadina and glutenins which play main role in determining of technological quality properties of wheat seeds and wheat products, but may be cause of allergy and celiac diseases in imune sensitive people. In this analysis of 10 wheat genotypes were established differences for gluten content, protein content loaf volume in two year of studies under different conditions. The highest gluten content in first year had Aurora (32.40%) and the lowest had G-3089 (21.40%), while in second year the highest gluten content had Beogradjanka (33.18%) and the lowest (22.33%) was in G-3089. In both year of experiment, the genotype G-3617 had the highest protein content (14.210%; 14.80%), loaf volume (560ml; 560ml) while genotypes G-3089 the lowest value of protein content (10.80%; 11.10%) and loaf volume (360ml; 380ml). In analysed wheat genotypes were identified 3 alleles (a, b, f) at Gli-A1 locus and 5 alleles (b, g, e, l, k) at Gli-A2 locus. Also, identified 3 alleles (a, b, c) at Glu-A1, 3 (a, c, d) at Glu-D1 and 5 alleles (a, b, c, d, h) at Glu-B1. The genotypes which possessed the alleles Glu-A1b, Glu-D1d, in combination Gli-A1b, had higher levels of gluten content, protein and bread volume.Gluten je kompleks glijadina i glutenina, koji ima značajnu ulogu u determinaciji osobina tehnološkog kvaliteta semena pšenice i gotovih proizvoda, a, može biti uzročnik alergijskih reakcija i nekih oboljenja probavnog trakta kod imunoosetljivih individua. U ovom radu je izučavano variranje sadržaja suvog glutena kod 10 genotipova pšenice, volumen hleba, sadržaj proteina, aleli glijadinskih alela sa 1A i 6A hromozoma i aleli sa visoko-molekularih glutenina na sva tri Glu-1 lokusa. Ustanovljeno je variranje: sadržaja suvog glutena u rasponu od 22,33% (G-3089) do 33,18% (Beogradjanka), sadržaja proteina od 10,80% (G-3089) do 14,80% (G-3617) i volumen hleba od 360ml (G-3089) do 560ml (G-3617). Na lokusu Gli-A1 identifikovano je 3 alela (a, b, f), na Gli-A2 je 5 alela (b, g, e, l, k). Determinisane su različite komponente visokomolekularnih glutenina i identifikovani aleli za te komponente. Ustanovljeno je prisistvo heterogenosti sorti na Glu-A1 kod dve sorte (Aurora i Kavkaz) i na Glu-D1 kod sorte Banaćanka 2. Identifikovano je po tri različita lalela na Glu-A1 (a, b, c) i Glu-D1 (a, c, d) i 5 alela na Glu-B1 (a, b ,c, d, h), a ustanovljeno je 9 tipova kompozicije gluteninskih alela

Analiza proteina u semenu pšenice (Triticum aestivum L.)

The protein in seed of wheat play a major role in determining the quality of wheat. The aim is to study the variability of the composition of the gene alleles of gliadins, and the glutenin content, total protein content and dry gluten in bread wheat genotypes. For this study were used seed of bread wheat genotypes grown in two years with different weather conditions. The results showed differences between varieties according to the composition of gliadin and glutenin, and according to the obtained values for the content of protein and gluten content. At the three Glu-1 loci were identified nine alleles, while at the three Gli-1 loci 13 alleles and at the three Gli-2 loci were identified 16 alleles. Composition of identified alleles was different in the wheat genotypes, and composition of glutenin alleles was specific for seven genotypes while three genotypes (G-3539/03, Poljana and G-3602/03) possessed the same allele (b, c, d). The protein and gluten content is varied depending on the genotype and environmental factors, The genotypes G-3602/03 had the lowest contents of protein 11,6% and gluten 10.018, while G-3539/03 had the highest content of protein 15.00% and dry gluten 14.086% in both years of experiment. Varying the content and composition of proteins is correlated with the properties of wheat quality.Proteini u semenu pšenice imaju glavnu ulogu u formiranju kvaliteta pšenice. Cilj rada je izučavanje varijabilnosti kompozicije genskih alela za glijadine i glutenine, sadržaja ukupnih proteina i sadržaja suvog glutena kod genotipova hlebne pšenice. Za ovo izučavanje je korišćeno seme 10 genotipova hlebne pšenice gajene u dve godine sa različitim vremenskim uslovima. Rezultati istraživanja su pokazali razlike izmedju sorti prema kompoziciji glijadina i glutenina, kao i prema dobijenim vrednostima za sadržaj proteina i sadržaj glutena. Na tri lokusa za visokomolekularne glutenine (Glu-1) je identifikovano devet alela, a na šest lokusa za glijadine je identifikovano ukupno 29 alela i to 13 alela na tri lokusa Gli-1 i 16 alela na tri lokusa Gli-2. Sastav glijadinskih alela je bio različit kod izučavanih genotipova pšenice, dok je sastav gluteninskih alela bio isti (b, c, d) kod tri genotipa (G-3539/03, Poljana i G-3602/03) a kod ostalih sedam sorti je bio različit. Sadržaj proteina je varirao zavisno od genotipa i od godine eksperimenta i to od 11,60% (G-3602/03 u prvoj godini) do 15,00% (G-3539/03 u drugoj godini). Sadržaj suvog glutena je varirao od 10,018% (G-3602/03 u prvoj godini) do 14,0860% (G-3539/03 u drugoj godini istraživanja). Kod izučavanih genotipova pšenice je ustanovljena heterogenost Glu-A1 lokusa (kod G-3908/03) i Glu-B1 lokusa (G-3602/03), kao i heterogenost Gli-A1 lokusa (G-3504/03 i G-3532/03), Gli-D1 lokusa (G-3908/03) i Gli-A2 lokusa (G-3908/03 i Žitnica). Kompozicija alela je bila ista u obe godine kod svih 10 genotipova, a sadržaj proteina i glutena je varirao, čije vrednosti su bile veće u drugoj godini eksperimenta

Effects of environment on optimizing nitrogen nutrition on content of first-class seeds in winter barley cultivars

In this paper was investigated variability of percentage content of seeds first-class (thickness of seeds >2.5mm) in four divergent barley cultivars (Jagodinac, Premium, NS 489 and NS 495) originated from different breeding centres in Serbia. These cultivars investigated during two years on experimental field on plots 5m2 and four replications under four different rate of nitrogen treatments (control N0=0, N1=20, N2=40 and N3=60 kg ha-1). The variability of content of seeds first-class was established in both years of experiment and under all nitrogen rate of supplied N fertilizer. According to percentage of content first-class (thickness of seeds >2.5mm) were found differences among the cultivars in each years of experiment and all variant of nitrogen nutrition. In average, content of seeds first-class for all cultivars and all treatments was higher in second year 82.34% than in first experimental year 80.03%. The average value of percentage content of seeds first-class in both growing seasons and under N treatment was the highest in barley cultivar NS 595 (82.70%) and the lowest in barley cultivar Premium (78.79%). For all investigated cultivars, content of seeds first-class increased with increasing nitrogen rate. This indicate that content of seeds first-class mostly affected by mineral nitrogen nutrition and to a much lesser degree by cultivar

Germination of maize hybrids seed stored after harvest

In this investigation used three maize hybrids that are belongs to different group of ripening: FAO 3, FAO 4 and FAO 6. The seed of those hybrids used for investigation of genotypic divergences for seed germination, energy of germination, content of abnormal emergence of seeds, content of dead seeds. Hybrid seed for analysis produced on the experimental field of the Institute of Maize in Zemun Polje, Belgrade. The viability of the maize hybrids seeds were determined in experimental analysis carried out in laboratory for quality of seed in the Maize Research Institute Zemun Polje, Belgrade. According to standard method on filter paper were estimated the values of seed germination. The three experiment of seed germination carried out: 2 months after harvest, 9 months after harvest and 18 months after harvest. In average for each test the highest percentage of seed germination were found for hybrid ZP FAO3: 92.75% for seed after 2 months of harvest, 92.00% for seed 9 monhts after harvest and 91.50% for seeds after 18 months of harvest. This hybrid had the lowest percentage of ungerminated seeds (2.75%) and the least content of dead seed (~5.00%). The lowest value of seed germination (86.33%) and the highest values of ungerminated seed (~3.00%) as well the highest content of dead seed (10.33%) had hybrid belongs FAO 4 group. The obtained results indicate differences of germination in analyzed maize genotypes

Germination of maize hybrids seed stored after harvest

In this investigation used three maize hybrids that are belongs to different group of ripening: FAO 3, FAO 4 and FAO 6. The seed of those hybrids used for investigation of genotypic divergences for seed germination, energy of germination, content of abnormal emergence of seeds, content of dead seeds. Hybrid seed for analysis produced on the experimental field of the Institute of Maize in Zemun Polje, Belgrade. The viability of the maize hybrids seeds were determined in experimental analysis carried out in laboratory for quality of seed in the Maize Research Institute Zemun Polje, Belgrade. According to standard method on filter paper were estimated the values of seed germination. The three experiment of seed germination carried out: 2 months after harvest, 9 months after harvest and 18 months after harvest. In average for each test the highest percentage of seed germination were found for hybrid ZP FAO3: 92.75% for seed after 2 months of harvest, 92.00% for seed 9 monhts after harvest and 91.50% for seeds after 18 months of harvest. This hybrid had the lowest percentage of ungerminated seeds (2.75%) and the least content of dead seed (~5.00%). The lowest value of seed germination (86.33%) and the highest values of ungerminated seed (~3.00%) as well the highest content of dead seed (10.33%) had hybrid belongs FAO 4 group. The obtained results indicate differences of germination in analyzed maize genotypes

Targeting MLL Methyltransferases Enhances the Antitumor Effects of PI3K Inhibition in Hormone Receptor–positive Breast Cancer

The high frequency of aberrant PI3K pathway activation in hormone receptor–positive (HR⁺) breast cancer has led to the development, clinical testing, and approval of the p110α-selective PI3K inhibitor alpelisib. The limited clinical efficacy of alpelisib and other PI3K inhibitors is partially attributed to the functional antagonism between PI3K and estrogen receptor (ER) signaling, which is mitigated via combined PI3K inhibition and endocrine therapy. We and others have previously demonstrated chromatin-associated mechanisms by which PI3K supports cancer development and antagonizes ER signaling through the modulation of the H3K4 methylation axis, inhibition of KDM5A promoter H3K4 demethylation and KMT2D/MLL4-directed enhancer H3K4 methylation. Here we show that inhibition of the H3K4 histone methyltransferase MLL1 in combination with PI3K inhibition impairs HR+ breast cancer clonogenicity and cell proliferation. While combined PI3K/MLL1 inhibition reduces PI3K/AKT signaling and H3K4 methylation, MLL1 inhibition increases PI3K/AKT signaling through the dysregulation of gene expression associated with AKT activation. These data reveal a feedback loop between MLL1 and AKT whereby MLL1 inhibition reactivates AKT. We show that combined PI3K and MLL1 inhibition synergizes to cause cell death in in vitro and in vivo models of HR⁺ breast cancer, which is enhanced by the additional genetic ablation of the H3K4 methyltransferase and AKT target KMT2D/MLL4. Together, our data provide evidence of a feedback mechanism connecting histone methylation with AKT and may support the preclinical development and testing of pan-MLL inhibitors. Significance: Here the authors leverage PI3K/AKT-driven chromatin modification to identify histone methyltransferases as a therapeutic target. Dual PI3K and MLL inhibition synergize to reduce clonogenicity and cell proliferation, and promote in vivo tumor regression. These findings suggest patients with PIK3CA-mutant, HR⁺ breast cancer may derive clinical benefit from combined PI3K/MLL inhibition.ISSN:2767-976