A Novel Transmembrane Semaphorin Can Bind c-src

doi:10.1006/mcne.1997.0644

Molecular and Cellular Neuroscience

Volume 9, Issues 5–6, 1997, Pages 409-419

https://doi.org/10.1006/mcne.1997.0644 Get rights and content

Abstract

The semaphorins/collapsins constitute a family of genes unified by the presence of a “semaphorin domain” which has been conserved through metazoan evolution. The semaphorin family comprises both secreted and transmembrane molecules and is thought to be made up of ligands for as yet unidentified receptors. The functions are not known, with the exception of those of sema III (also referred as sem D and collapsin 1), D-sema I, and D-sema II, which have been shown to be involved in axonal pathfinding. Here we report the identification of a mouse semaphorin cDNA, termedSema VIb.Although Sema VIb contains the extracellular semaphorin domain, it lacks the immunoglobulin domain or thrombospondin repeats which are present in other described vertebrate (but not invertebrate) transmembrane semaphorins. During developmentSema VIbmRNA is expressed in subregions of the nervous system and is particularly prominent in muscle. In adulthood,Sema VIbmRNA is expressed ubiquitously. The cytoplasmic domain of Sema VIb contains several proline-rich potential SH3 domain binding sites. Using anin vitrobinding assay, we show that Sema VIb binds specifically the SH3 domain of the protooncogene c-src. In transfected COS cells Sema VIb coimmunoprecipitates with c-src. These results, along with our evidence that Sema VIb can form dimers, suggests that the semaphorin family not only serves as ligands but may include members, especially those which are transmembrane, which serve as receptors, triggering intracellular signaling via an src-related cascade.

References (35)

R.H. Adams et al.
A novel class of murine semaphorins with homology to thrombospondin is differentially expressed during early embrygenesis
Mech. Dev.
(1996)
G.B. Cohen et al.
Modular binding domains in signal transduction proteins
Cell
(1995)
A.L. Kolodkin et al.
The semaphorin genes encode a family of transmembrane and secreted growth cone guidance molecules
Cell
(1993)
J. Kyte et al.
A simple method for displaying the hydropathic character of a protein
J. Mol. Biol.
(1982)
Y. Luo et al.
Collapsin: A protein in brain that induces the collapse and paralysis of neuronal growth cones
Cell
(1993)
Y. Luo et al.
A family of molecules related to collapsin in the embryonic chick nervous system
Neuron
(1995)
D.J. Matthes et al.
Semaphorin II can function as a selective inhibitor of specific synaptic arborizations
Cell
(1995)
E.K. Messersmith et al.
Semaphorin III can function as a selective chemorepellent to pattern sensory projections in the spinal cord
Neuron
(1995)
H. Yu et al.
Structural basis for the binding of proline-rich peptides to SH3 domains
Cell
(1994)
L. Zhou et al.
Cloning and expression of a novel murine semaphorin with structural similarity to insect semaphorin I
Mol. Cell. Neurosci.
(1997)

F.M. Ausubel et al.

Current Protocols in Molecular Biology

(1989)

O. Behar et al.

Semaphorin III is needed for normal patterning and growth of nerves, bones and heart

Nature

(1996)

J.K. Chen et al.

Biased combinatorial libraries: Novel ligands for the SH3 domain of phosphatidylinositol 3-kinase

J. Am. Chem. Soc.

(1993)

P. Cicchetti et al.

Identification of a protein that binds to the SH3 region of Abl and is similar to Bcr and GAP-rho

Science

(1992)

D.G. Drubin et al.

Homology of a yeast actin-binding protein to signal transduction proteins and myosin-I

Nature

(1990)

T. Erpel et al.

Mutational analysis of the SRC SH3 domain: The same residues of the ligand binding surface are important for intra- and intermolecular interactions

EMBO J.

(1995)

G.I. Evan et al.

Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product

Mol. Cell. Biol.

(1985)

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The semaphorin protein family controls a wide variety of developmental processes in the nervous system such as axon guidance, cell migration, dendrite morphology, and synaptogenesis. The best characterized function of semaphorins is their ability to attract or repel neurites, directing these structures toward or away from specific regions. Although only a limited set of semaphorins has been identified, accumulating evidence indicates that distinct molecular mechanisms act to diversify the effects of these guidance cues, allowing them to control a disproportionally large number of different cellular events. In this chapter, we discuss general features and novel aspects of semaphorin function and signaling, including the role of their receptors: plexins and neuropilins. The focus is on the contribution of these molecules to the regulation of cell migration and the formation of neuronal connections. In addition, we highlight a few recent studies that examine how semaphorins, plexins, and/or neuropilins contribute to neurological diseases.
Analysis of SEMA6B gene expression in breast cancer: Identification of a new isoform
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Citation Excerpt :
Such semaphorins can be further processed into soluble forms through proteolytic degradation, as observed for SEMA4D [10,11]. Membrane-bound semaphorins that do not appear to be processed in soluble forms, such as semaphorin 6 (from A to D), are involved in reverse signaling mechanisms [12–14]. The human SEMA6B gene was first identified in 2001 by EST sequencing from a cDNA pool of breast cancer donors, and its expression was described in MCF-7 cell line, a widely used breast cancer-derived cell line [15].
SEMA6B is a member of the semaphorins axon-guidance family. A growing body of evidence has been accumulated describing the role of semaphorin molecules in cancer development and the involvement of SEMA6B in cancer progression has recently been proposed.
Our analysis, based on real-time PCR, focused on the expression of SEMA6B in a panel of breast cancer tissues, compared to the normal counterpart.
In cancer tissues we found a significantly strong down-modulation of this transcript. Moreover we identified and characterized a novel SEMA6B isoform, named SEMA6Ba. This isoform has a novel splice junction, created by the usage of alternative donor and acceptor splice sites internal to the exon 17. By in silico analysis we found that the new transcript 3′ UTR lacks some highly-conserved miRNA binding sites, suggesting possible consequences on both spatial and temporal expression of SEMA6Ba. The translated sequence of SEMA6Ba lacks the cytoplasmic tail, crucial for triggering the reverse signaling described for the transmembrane semaphorins. We also demonstrated, by immunofluorescence analysis of endogenous and overexpressed SEMA6Ba, that the protein clearly localized to the endoplasmic reticulum and plasma membrane. In conclusion, SEMA6B gene products are strongly down modulated in breast cancer tissues and a new isoform named SEMA6Ba has been described and characterized.
Our work states a clear relation among breast cancer and SEMA6B expression; moreover we describe for the first time the SEMA6Ba protein and report here the analysis of SEMA6Ba RNA messenger, the protein expression and the cellular localization.
Semaphorins and their Receptors in Vertebrates and Invertebrates
2009, Handbook of Cell Signaling, Second Edition
Semaphorins are a large family of proteins originally identified as axon guidance factors of the developing nervous system. Semaphorins act as both repellents and attractants for growing axons and thus have a significant function in central nervous system (CNS) development. Semaphorins may guide growing dendrites as well as axons. Many types of neurons are responsive to semaphorins, including dorsal root ganglion, sensory, motor, hippocampal, cortical, cerebellar, and olfactory. In addition to guiding axons and dendrites, semaphorins appear to play a role in fasciculation of nerve bundles, neuronal cell migration, axoplasmic transport, and apoptosis. Apart from the CNS, migrating non-neuronal cells are responsive to semaphorins, and cardiovascular abnormalities are observed when semaphorin signaling is disrupted. In the immune system, expression of Sema4D (CD100) is regulated upon B- and T-lymphocyte activation, and migrating monocytes are responsive to Sema3A and Sema4D. Malignant lung cells show reduced levels and a cytoplasmic localization of semaphorins. Therefore, it can be concluded that semaphorins act as guidance cues for many types of migrating cells in developmental, adult, and pathological tissue. Thus, many biological systems in the developing embryo and adult animal are dependent on semaphorin signaling. Although the importance of semaphorins in the developing CNS is well documented, their involvement in the immune response, the cardiovascular system, and in pathology is still being clarified.
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2008, Trends in Biochemical Sciences
Semaphorins were initially characterized according to their role in repulsive axon guidance but are now recognized as crucial regulators of morphogenesis and homeostasis over a wide range of organ systems. The pleiotropic nature of semaphorin signaling and its implication in human disease has triggered an enormous interest in the receptor and intracellular signaling mechanisms that direct the cell-type-specific and diverse biological effects of semaphorins. Recent breakthroughs in our understanding of semaphorin signaling link integrin and semaphorin signaling pathways, identify novel ligand–receptor interactions and provide insight into the cellular and molecular bases of bifunctional and reverse signaling events. These discoveries could lead to therapeutic advances in axonal regeneration, cancer and other diseases.
Semaphorins in development and adult brain: Implication for neurological diseases
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As a group, Semaphorins are expressed in most tissues and this distribution varies considerably with age. Semaphorins are dynamically expressed during embryonic development and their expression is often associated with growing axons. This expression decreases with maturity and several observations support the idea that in adult brain the expression of secreted Semaphorins is sensitive to electrical activity and experience. The functional role of Semaphorins in guiding axonal projections is well established and more recent evidence points to additional roles in the development, function and reorganization of synaptic complexes. Semaphorins exert the majority of their effects by binding to cognate receptor proteins through their extracellular domains. A common theme is that Semaphorin-triggered signalling induces the rearrangement of the actin and microtubule cytoskeleton. Mutations in Semaphorin genes are linked to several human diseases associated with neurological changes, but their actual influence in the pathogenesis of these diseases remains to be demonstrated. In addition, Semaphorins and their receptors are likely to mediate cross-talk between neurons and other cell types, including in pathological situations where their influence can be damaging or favourable depending on the context. We discuss how the manipulation of Semaphorin function might be crucial for future clinical studies.
Graded Expression of Semaphorin-1a Cell-Autonomously Directs Dendritic Targeting of Olfactory Projection Neurons
2007, Cell
Citation Excerpt :
Do transmembrane semaphorins also function cell-autonomously as axon guidance receptors in the nervous system? This possibility has been proposed in a number of reports (Eckhardt et al., 1997; Klostermann et al., 2000; Leighton et al., 2001; Godenschwege et al., 2002), and was shown to be the case for Drosophila Sema-1a in photoreceptor axon targeting (Cafferty et al., 2006). Below we provide evidence that Sema-1a functions as a receptor in axon targeting of PNs and mushroom body (MB) neurons.
Gradients of axon guidance molecules instruct the formation of continuous neural maps, such as the retinotopic map in the vertebrate visual system. Here we show that molecular gradients can also instruct the formation of a discrete neural map. In the fly olfactory system, axons of 50 classes of olfactory receptor neurons (ORNs) and dendrites of 50 classes of projection neurons (PNs) form one-to-one connections at discrete units called glomeruli. We provide expression, loss- and gain-of-function data to demonstrate that the levels of transmembrane Semaphorin-1a (Sema-1a), acting cell-autonomously as a receptor or part of a receptor complex, direct the dendritic targeting of PNs along the dorsolateral to ventromedial axis of the antennal lobe. Sema-1a also regulates PN axon targeting in higher olfactory centers. Thus, graded expression of Sema-1a contributes to connection specificity from ORNs to PNs and then to higher brain centers, ensuring proper representation of olfactory information in the brain.

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Regular ArticleA Novel Transmembrane Semaphorin Can Bind c-src

Abstract

Mech. Dev.

Cell

Cell

J. Mol. Biol.

Cell

Neuron

Cell

Neuron

Cell

Mol. Cell. Neurosci.

Current Protocols in Molecular Biology

Semaphorin III is needed for normal patterning and growth of nerves, bones and heart

Nature

Biased combinatorial libraries: Novel ligands for the SH3 domain of phosphatidylinositol 3-kinase

J. Am. Chem. Soc.

Identification of a protein that binds to the SH3 region of Abl and is similar to Bcr and GAP-rho

Science

Homology of a yeast actin-binding protein to signal transduction proteins and myosin-I

Nature

Mutational analysis of the SRC SH3 domain: The same residues of the ligand binding surface are important for intra- and intermolecular interactions

EMBO J.

Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product

Mol. Cell. Biol.

Regular Article
A Novel Transmembrane Semaphorin Can Bind c-src