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  • 1.
    Gallio, Marco
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap. Karolinska Institutet.
    The Rhomboid family of intramembrane proteases, conserved regulators of cell communication2004Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    The development of multicellular organisms relies heavily on cell communication. Cells send and receive complex sets of signals, harmonising their growth and differentiation with that of other, often distant, cell populations. In animals, the Epidermal Growth Factor Receptor (EGFR) is an important mediator of cell communication. EGFR activation regulates various developmental events in nematodes, insects and vertebrates. In addition, mutations in human EGFRs have been associated with a number of cancers. In Drosophila, a key event triggering EGFR signalling is the regulated release of the extracellular portion of EGFR ligands. Rhomboid (Rho), an unusual polytopic protease, cleaves the transmembrane, inactive ligand precursor into an active, soluble form. Both the target sequence and Rho s catalytic site are embedded within the membrane bilayer and for this reason the reaction has been described as regulated intramembrane proteolysis. The work presented in this thesis begins with the characterisation of a classical fly mutation, roughoid (ru). Our results indicate that ru acts as a novel, positive regulator of EGFR signalling during eye development in Drosophila. ru was subsequently identified as rhomboid-3, one of seven rhomboid related genes encoded in the fly genome. Unexpectedly, we found that sequences related to Rhomboid are also common in unicellular organisms. A single microbial Rho has been previously studied, the aarA gene from the human pathogen Providencia stuartii. Strikingly, AarA appears to have a corresponding function to that of the Drosophila Rho: it is necessary for the release of a peptide-signal, which mediates cell communication in P. stuartii. AarA was indeed capable of substituting for the fly Rho in vivo. Vice versa, the fly Rho-1 restored the ability of aarA mutant bacteria to produce the extracellular signal mediating cell communication. These results suggest that Rho-mediated proteolysis might represent a very ancient mechanism for cell communication. The Drosophila genome contains seven Rhomboids. We began to investigate the possibility of additional substrates by analyzing the respiratory system phenotype observed in ru/rho-3 mutant embryos. During embryogenesis, specialised tracheal branches target and invade the ventral nerve cord, part of the central nervous system (CNS). In ru/rho-3 mutants, these branches are misrouted, and inappropriately cross the CNS midline. Also in this context Rho-3 functions to activate an EGFR ligand. Yet, the results reveal an unusual role for the pathway in the repulsion of migrating epithelial cells. EGFR ligands act as chemoattractants for a variety of cells in vivo and in vitro, including tumors. Our results provide a proof of principle that the EGFR can also mediate repulsion from the signal source.

  • 2.
    Gallio, Marco
    et al.
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap. Stockholm University / Karolinska Institute.
    Englund, C
    Stockholm University / Umeå University.
    Kylsten, Per
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap.
    Samakovlis, C
    Stockholm University.
    Rhomboid 3 orchestrates Slit-independent repulsion of tracheal branches at the CNS midline2004Ingår i: Development, ISSN 0950-1991, E-ISSN 1477-9129, Vol. 131, nr 15, s. 3605-3614Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    EGF-receptor ligands act as chemoattractants for migrating epithelial cells during organogenesis and wound healing. We present evidence that Rhomboid 3/EGF signalling, which originates from the midline of the Drosophila ventral nerve cord, repels tracheal ganglionic branches and prevents them from crossing it. rho3 acts independently from the main midline repellent Slit, and originates from a different sub-population of midline cells: the VUM neurons. Expression of dominant-negative Egfr or Ras induces midline crosses, whereas activation of the Egfr or Ras in the leading cell of the ganglionic branch can induce premature turns away from the midline. This suggests that the level of Egfr intracellular signalling, rather than the asymmetric activation of the receptor on the cell surface, is an important determinant in ganglionic branch repulsion. We propose that Egfr activation provides a necessary switch for the interpretation of a yet unknown repellent function of the midline.

  • 3.
    Gallio, Marco
    et al.
    Södertörns högskola, Avdelning Naturvetenskap. Karolinska Institutet.
    Kylsten, Per
    Södertörns högskola, Avdelning Naturvetenskap. Karolinska Institutet.
    Providencia may help find a function for a novel, widespread protein family2000Ingår i: Current Biology, ISSN 0960-9822, E-ISSN 1879-0445, Vol. 10, nr 19, s. R693-R694Artikel i tidskrift (Refereegranskat)
  • 4.
    Gallio, Marco
    et al.
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap. Karolinska Institutet.
    Kylsten, Per
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap.
    The roughoid locus identifies a novel function involved in epidermal growth factor receptor signalling in DrosophilaManuskript (preprint) (Övrigt vetenskapligt)
  • 5.
    Gallio, Marco
    et al.
    Södertörns högskola, Avdelning Naturvetenskap. Karolinska Institute.
    Sturgill, G
    Case Western Reserve University School of Medicine, Cleveland, USA.
    Rather, P
    Case Western Reserve University School of Medicine, Cleveland, USA.
    Kylsten, Per
    Södertörns högskola, Avdelning Naturvetenskap.
    A conserved mechanism for extracellular signaling in eukaryotes and prokaryotes2002Ingår i: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 99, nr 19, s. 12208-12213Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Epidermal growth factor receptor (EGFr) is a key mediator of cell communication during animal development and homeostasis. In Drosophila, the signaling event is commonly regulated by the polytopic membrane protein Rhomboid (RHO), which mediates the proteolytic activation of EGFr ligands, allowing the secretion of the active signal. Until very recently, the biochemical function of RHO had remained elusive. It is now believed that Drosophila RHO is the founder member of a previously undescribed family of serine proteases, and that it could be directly responsible for the unusual, intramembranous cleavage of EGFr ligands. Here we show that the function of RHO is conserved in Gram-negative bacteria. AarA, a Providencia stuartii RHO-related protein, is active in Drosophila on the fly EGFr ligands. Vice versa, Drosophila RHO-1 can effectively rescue the bacterium's ability to produce or release the signal that activates density-dependent gene regulation (or quorum sensing). This study provides the first evidence that prokaryotic and eukaryotic RHOs could have a conserved role in cell communication and that their biochemical properties could be more similar than previously anticipated.

  • 6. Lundström, A
    et al.
    Gallio, Marco
    Södertörns högskola, Institutionen för kemi, biologi, geografi och miljövetenskap.
    Englund, C
    Steneberg, P
    Hemphälä, J
    Aspenström, P
    Keleman, K
    Falileeva, L
    Dickson, B J
    Samakovlis, C
    Vilse, a conserved Rac/Cdc42 GAP mediating Robo repulsion in tracheal cells and axons2004Ingår i: Genes & Development, ISSN 0890-9369, E-ISSN 1549-5477, Vol. 18, nr 17, s. 2161-2171Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Slit proteins steer the migration of many cell types through their binding to Robo receptors, but how Robo controls cell motility is not clear. We describe the functional analysis of vilse, a Drosophila gene required for Robo repulsion in epithelial cells and axons. Vilse defines a conserved family of RhoGAPs (Rho GTPase-activating proteins), with representatives in flies and vertebrates. The phenotypes of vilse mutants resemble the tracheal and axonal phenotypes of Slit and Robo mutants at the CNS midline. Dosage-sensitive genetic interactions between vilse, slit, and robo mutants suggest that vilse is a component of robo signaling. Moreover, overexpression of Vilse in the trachea of robo mutants ameliorates the phenotypes of robo, indicating that Vilse acts downstream of Robo to mediate midline repulsion. Vilse and its human homolog bind directly to the intracellular domains of the corresponding Robo receptors and promote the hydrolysis of RacGTP and, less efficiently, of Cdc42GTP. These results together with genetic interaction experiments with robo, vilse, and rac mutants suggest a mechanism whereby Robo repulsion is mediated by the localized inactivation of Rac through Vilse.

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