Cloning and characterization of a novel epidermal cell surface antigen (ESA).

Schroeder, W. T.; Stewart-Galetka, Shelley; Mandavilli, Srinivas; Goldsmith, Lowell A.; Duvic, Madeleine

doi:10.1016/s0021-9258(17)32117-8

Cited by 37 publications

(11 citation statements)

References 41 publications

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“…Degenerate sense and antisense primers were designed for PCR on the basis of multiple sequence alignments of goldfish reggie-1 (Schulte et al, 1997), ESA, and MESA cDNAs (Schroeder et al, 1994), and of goldfish reggie-2 (Schulte et al, 1997) and human EST clones (HO3390, H14248, and H30476), respectively. The template for PCR was oligo(dT)-primed first-strand cDNA reverse-transcribed from 2 g poly(A) ϩ RNA using Superscript II reverse transcriptase (Gibco BRL Life Technologies, Eggenstein, Germany).…”

Section: Cloning Of Partial Rat Reggie-1 and Reggie-2 Cdnasmentioning

confidence: 99%

“…An RT-PCR cloning strategy with degenerate oligonucleotide primers designed on the basis of sequence comparisons of goldfish reggie-1 with ESA/ MESA, and goldfish reggie-2 (Schulte et al, 1997;Schroeder et al, 1994) with human EST clones was employed to isolate partial rat reggie-1 and rat reggie-2 cDNAs. The resulting PCR fragments were used to screen a cDNA library of 14-to 16-day-old rat brains.…”

Section: Molecular Cloning Of Rat Reggie-1 and -2mentioning

confidence: 99%

“…A 47-kD protein, flotillin-1, has been identified in a nonionic detergent insoluble fraction from the brain (Bickel et al, 1997). Flotillin-1 and a protein with significant sequence homologies named "epidermal surface antigen" (ESA) or (mouse) MESA (Schroeder et al, 1994) were suggested to represent resident components of caveolae in fibroblasts and lung endothelial cells and to be associated with caveolae or caveolae-like structures in neural tissue (Bickel et al, 1997).…”

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confidence: 99%

“…Mouse flotillin-1 is the homolog of a protein which we discovered independently in the goldfish central nervous system (CNS) and which we named "reggie-2" (Schulte et al, 1997) because the mRNAs of reggie-2 and a 44% homologous protein, reggie-1, are up-regulated in fish retinal ganglion cells (RGCs) after optic nerve transection and during RGC axon regeneration. In overlapping segments, goldfish reg-gie-1 is 80% identical with ESA/MESA, which was described as a 35-kD surface protein in keratinocytes of mice and humans (Schroeder et al, 1994).…”

mentioning

confidence: 99%

“…Moreover, to functionally characterize reggie-1 and -2 and to determine whether they form microdomains where activated GPI-linked proteins cluster, the use of mammalian neurons and cell lines seemed advantageous because many more relevant proteins are known and antibodies are available to perform such experiments. When these experiments were initiated, sequence information on reggie-1 and -2 in the rat brain was not yet available except for the sequence of ESA/MESA, claimed to represent a cell-surface protein of human and mouse keratinocytes (Schroeder et al, 1994). Moreover, cRNA probes derived from the fish cDNA did not hybridize with the presumed rat reggie-1 and -2 mRNAs.…”

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confidence: 99%

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Identification of Reggie-1 and Reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons

et al. 1998

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Neurons are believed to possess plasmalemmal microdomains and proteins analogous to the caveolae and caveolin of nonneuronal cells. Caveolae are plasmalemmal invaginations where activated glycosyl‐phosphatidylinositol (GPI)‐anchored proteins preferentially assemble and where transmembrane signaling may occur. Molecular cloning of rat reggie‐1 and ‐2 (80% identical to goldfish reggie proteins) shows that reggie‐2 is practically identical to mouse flotillin‐1. Flotillin‐1 and epidermal surface antigen (ESA) (flotillin‐2) are suggested to represent possible membrane proteins in caveolae. Rat reggie‐1 is 99% homologous to ESA in overlapping sequences but has a 49‐amino‐acid N‐terminus not present in ESA. Antibodies (ABs) which recognize reggie‐1 or ‐2 reveal that both proteins cluster at the plasmamembrane and occur in micropatches in neurons [dorsal root ganglia (DRGs), retinal ganglion, and PC‐12 cells] and in nonneuronal cells. In neurons, reggie micropatches occur along the axon and in lamellipodia and filopodia of growth cones, but they do not occur in caveolae. By quantitative electronmicroscopic analysis we demonstrate the absence of caveolae in (anti‐caveolin negative) neurons and show anti‐reggie‐1 immunogold‐labeled clusters at the plasmamembrane of DRGs. When ABs against the GPI‐anchored cell adhesion molecules (CAMs) F3 and Thy‐1 are applied to live DRGs, the GPI‐linked CAMs sequester into micropatches. Double immunofluorescence shows a colocalization of the CAMs with micropatches of anti‐reggie antibodies. Thus, reggie‐1 and reggie‐2 identify sites where activated GPI‐linked CAMs preferentially accumulate and which may represent noncaveolar micropatches (domains). © 1998 John Wiley & Sons, Inc. J Neurobiol 37: 502–523, 1998

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Section: Cloning Of Partial Rat Reggie-1 and Reggie-2 Cdnasmentioning

confidence: 99%

Section: Molecular Cloning Of Rat Reggie-1 and -2mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Identification of Reggie-1 and Reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons

et al. 1998

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Flotillin 2 is distinct from epidermal surface antigen (ESA) and is associated with filopodia formation

Hazarika¹,

Dham²,

Patel³

et al. 1999

J. Cell. Biochem.

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ECS-1, a monoclonal antibody (MoAb) raised to cultured human keratinocytes, stains the intercellular glycocalyx with a pemphigus-like pattern and recognizes a 35-kDa epidermal surface antigen (ESA) on Western blotting of keratinocyte extracts. When ECS-1 MoAb was used to screen a keratinocyte expression library, a unique cDNA was identified that predicted a 42-kDa globular protein of unknown function. This putative ESA was conserved between mice and humans and was encoded by a gene on chromosome 17q11-12 in linkage with neurofibromin. Homology between the cDNA sequence has been reported with flotillin 1, a caveolae associated protein, as well as Reggie 1 and 2, neuronal proteins expressed during axonal regeneration present in activated GPI-anchored cell adhesion molecules in non-caveolar-associated micropatches. In order to determine whether the cDNA predicted protein and ECS-1 antigen were identical, we compared ECS-1 with the immunoreactivity of a new antibody raised to the cDNA fusion protein in epidermis and cultured cells. The cDNA fusion protein was expressed in bacteria and in cos cells with his, FLAG, and EGFP reporter tags and by stable transfection as an EGFP fusion protein. The fusion protein and native protein of 42 kDa were detected by the new antibody, but not by the original ECS-1. Thus, the ECS-1 antigen, ESA (35 kDa), is clearly distinct from the protein predicted by the cDNA (renamed flotillin 2). Stable transfection of ESA/flotillin 2 fusion protein in cos cells induced filopodia formation and changed epithelial cells to a neuronal appearance. Thus, the function of flotillin 2 may resemble that of the goldfish optic nerve neuronal regeneration proteins, Reggie 1 and 2.

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Restricted expression ofreggiegenes and proteins during early zebrafish development

et al. 2004

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Reggies are plasma membrane-associated proteins and characteristic markers of lipidraft microdomains. They are highly conserved from flies to humans and have been implicated in axon regeneration and cell process and contact formation, possibly providing functional platforms for cell-signaling in neurons and other cell types. We analyzed reggie mRNA and protein expression patterns during early zebrafish development. All three zebrafish genes, re-1a, -2a, and -2b, span a considerably diverse set of expression patterns, and their proteins are induced maternally, showing ubiquitous expression at early stages. Although re-2a mRNA can be observed in differentiating neurons in the brain, spinal cord, and neurogenic placodes, re-2b is transcribed mainly in head mesoderm, in neural crest derivates, and along somite boundaries. re-1a mRNA is present at high levels in expression domains that overlap with the combined expression pattern of both re-2 genes except at the somites, where it complements the pattern of re-2b. Immunostaining on embryos reveals reggie protein localization at the cell membrane, at cell-cell contacts, and along all early axon tracts. The early phase of reggie expression suggests a basic and ubiquitous function during the first stages of embryogenesis and into the gastrula period. Upon segmentation, a second phase of expression shows distinctly localized expression patterns, indicating tissue-specific roles and an involvement of re-1a/re-2a in neural development.

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Cloning and characterization of a novel epidermal cell surface antigen (ESA).

Cited by 37 publications

References 41 publications

Identification of Reggie-1 and Reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons

Identification of Reggie-1 and Reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons

Flotillin 2 is distinct from epidermal surface antigen (ESA) and is associated with filopodia formation

Restricted expression ofreggiegenes and proteins during early zebrafish development

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