ABSTRACT We generated Dictyostelium double mutants lacking the two F-actin crosslinking proteins α-actinin and gelation factor by inactivating corresponding genes via homologous recombination. Here we investigated consequences of these deficiencies both at single cell level multicellular stage. found that loss severely affected growth mutant cells in shaking suspension, led to a reduction size from 12 μm wild-type 9 cells. Moreover did not exhibit typical polarized morphology aggregating but...

Chemotaxis and phagocytosis are basically similar in cells of the immune system Dictyostelium amebae. Deletion unique G protein β subunit D. discoideum impaired but had little effect on fluid-phase endocytosis, cytokinesis, or random motility. Constitutive expression wild-type restored normal development. Chemoattractants released by bacteria trigger typical transient actin polymerization responses cells. In subunit–null cells, a series point mutants, these were to degree that correlated...

Vascular physiology relies on the concerted dynamics of several cell types, including pericytes, endothelial, and vascular smooth muscle cells. The interactions between such types are inherently dynamic not easily described with static, fixed, experimental approaches. Pericytes mural cells that support development, remodeling, homeostasis, involved in a number pathological situations cancer. interplay pericytes endothelial is at basis few tools exist to properly describe study it. Here we...

Dictyostelium amoebae are professional phagocytes, which ingest bacteria as the principal source of food. We have cloned homologue human natural resistance-associated membrane protein 1 (Nramp1) [solute carrier family 11 member (Slc11a1)], an endo-lysosomal that confers on macrophages resistance to infection by a variety intracellular and protozoa. The Nramp1 gene encodes 53 kDa with putative transmembrane domains. is transcribed during growth-phase downregulated barely detectable levels...

Abstract Background Phagocytosis plays a major role in the defense of higher organisms against microbial infection and provides also basis for antigen processing immune response. Cells model organism Dictyostelium are professional phagocytes that exploit phagocytosis bacteria as preferred way to ingest food, besides killing pathogens. We have investigated differential gene expression during non-pathogenic bacteria, using DNA microarrays, order identify molecular functions novel genes...

Membrane phosphatidylinositides recruit cytosolic proteins to regulate phagocytosis, macropinocytosis and endolysosomal vesicle maturation. Here, we describe effects of inactivation PI3K, PTEN or PLC on Escherichia coli Legionella pneumophila uptake by the professional phagocyte Dictyostelium discoideum. We show that L. is engulfed macropinocytosis, a process partially sensitive PI3K inactivation, unlike phagocytosis E. coli. Both processes are blocked inhibition. Whereas rapidly digested,...

The Dictyostelium discoideum genome harbors two genes encoding members of the Nramp superfamily, which is conserved from bacteria (MntH proteins) to humans (Slc11 proteins). Nramps are proton-driven metal ion transporters with a preference for iron and manganese. Acquisition these cations vital all cells, as they act redox cofactors regulate key cellular processes, such DNA synthesis, electron transport, energy metabolism oxidative stress. Nramp1 (Slc11a1), like its mammalian ortholog,...

Iron, zinc and copper play fundamental roles in eucaryotes procaryotes, their bioavailability regulates host-pathogen interactions. For intracellular pathogens, the source of metals is cytoplasm host, which turn manipulates metal traffic following pathogen recognition. It established that iron withheld from pathogen-containing vacuole, whereas for evidence unclear. Most infection studies mammals have concentrated on effects deficiency/overloading at organismal level. Thus, deficiency or...

The Nramp (Slc11) protein family is widespread in bacteria and eucaryotes, mediates transport of divalent metals across cellular membranes. social amoeba Dictyostelium discoideum harbours two proteins. Nramp1, like its mammalian ortholog, recruited to phagosomal macropinosomal membranes, confers resistance pathogenic bacteria. Nramp2 located exclusively the contractile vacuole membrane controls, synergistically with iron homeostasis. It has long been debated whether Nramp1 import or export...

The behavior of a cell is strongly influenced by the physical properties and stimuli in its microenvironment. Furthermore, activation modulation mechanotransduction pathways are involved tissue development homeostasis even pathological processes. Thus, when developing materials aimed at mimicking extracellular matrixes healthy or tissues, their mechanical features should be closely considered. In this context, nanoindentation represents powerful technique for mechanically characterizing...

// Valentina Monica 1 , Marco Lo Iacono Enrico Bracco Simone Busso Laura Di Blasio 2 Luca Primo Barbara Peracino 3 Mauro Papotti Giorgio Scagliotti Department of Oncology, University Torino, San Luigi Hospital, Orbassano, Italy IRCCS Candiolo, Clinical and Biological Sciences, Correspondence to: Monica, email: valentina.monica@unito.it Keywords: pemetrexed, dasatinib, malignant pleural mesothelioma, cell lines, thymidylate synthase Received: August 17, 2015 Accepted: June 16, 2016 Published:...

Metastasisation occurs through the acquisition of invasive and survival capabilities that allow tumour cells to colonise distant sites. While role multicellular aggregates in cancer dissemination is acknowledged, mechanisms drive formation multiclonal cell are not fully elucidated. Here, we show different tissue origins can perform collective directional migration actively form heteroclonal 3D, a proliferation-independent mechanism. Coalescence clusters mediated by subcellular actin-rich...

The main function of vacuolar H + ATPases in eukaryotic cells is to generate proton and electrochemical gradients across the membrane inner compartments. We have isolated gene encoding B subunit Dictyostelium discoideum ATPase ( vatB ) analyzed its transcriptional regulation. deduced protein comprises 493 amino acids with a calculated molecular mass 54 874 Da. predicted sequence highly homologous previously determined V/H sequences. encoded by single genome. maximally expressed during growth...

Recently we reported that Dictyostelium cells ingest Legionella pneumophila by macropinocytosis, whereas other bacteria, such as Escherichia coli, Mycobacterium avium, Neisseria meningitidis or Salmonella typhimurium, are taken up phagocytosis.1 In contrast to phagocytosis, macropinocytosis is partially inhibited PI3K PTEN inactivation, both processes sensitive PLC inhibition. Independently from reduced uptake, L. proliferates more efficiently in PI3K-null than wild-type cells. inactivation...

In Chronic Myeloid Leukemia 80% of patients present alternative splice variants involving BCR exons 1, 13 or 14 and ABL exon 4, with a consequent impairment in the reading frame gene. Therefore BCR/ABL fusion proteins (BCR/ABL-OOF) are characterized by an in-frame portion followed amino acids sequence arising from out (OOF) The product this new transcript contains characteristic domains while lacking COOH-terminal Rho GTPase GAP domain. work aims to characterize protein functionality terms...

Membrane proteins are involved in different physiological functions and the target of pharmaceutical abuse drugs. Xenopus laevis oocytes provide a powerful heterologous expression system for functional studies these proteins. Typical experiments investigate transport using electrophysiology radiolabeled uptake. A two-electrode voltage clamp is suitable only electrogenic proteins, uptake measurements require existence substrates adequate laboratory facilities.Recently, Dictyostelium...

Abstract Colorectal cancer (CRC) is one of the most diagnosed cancers worldwide. Though several treatment strategies are available today, around a million people die this disease each year Thus, there still pivotal need for new and more effective treatments. Genetic non-genetic intra-tumor heterogeneity has emerged lately as an intrinsic feature malignancies factors responsible therapy resistance or tolerance. In particular, nature role that phenotypically distinct, yet genetically...

ABSTRACT Metastasisation occurs through the acquisition of invasive and survival capabilities that allow tumour cells to colonise distant sites. While role multicellular aggregates in cancer dissemination is acknowledged, mechanisms drive formation multi-clonal cell are not fully elucidated. Here we show different tissue origins can perform collective directional migration actively form heteroclonal 3D, a proliferation-independent mechanism. Coalescence clusters mediated by subcellular...