Molecular cloning, expression and tissue distribution of glial-cell-line-derived neurotrophic factor family receptor alpha-3 (GFRalpha-3).
Neurturin and glial-cell-line-derived neurotrophic factor (GDNF) promote the survival and maintenance of different types of neuronal cells and signal through a receptor complex composed of a ligand binding subunit, either GDNF family receptor alpha-1 (GFRalpha-1) or alpha-2 (GFRalpha-2), together with the cRET membrane-bound protein tyrosine kinase. We have cloned GFRalpha-3, a novel receptor belonging to the GFRalpha family, that is 35% identical by amino acid sequence to both GFRalpha-1 and GFRalpha-2. GFRalpha-3 is a protein composed of 400 amino acid residues with three potential N-linked glycosylation sites together with the features characteristic of a glycosyl-phosphatidylinositol-anchored membrane protein. The heterologous expression of a FLAG-tagged GFRalpha-3 in human embryonic kidney cells showed that the protein is bound to the cell surface via a glycosyl-PtdIns anchor and is glycosylated, with different glycoforms migrating on SDS/PAGE with apparent molecular masses ranging over 43-62 kDa. The gene for GFRalpha-3 was mapped to human chromosome 5 in a region (q31.1-q31.3) where several disease loci, growth factor and growth factor receptor genes have been localized. Using northern blot analysis or reverse-transcription PCR, GFRalpha-3 was shown to be expressed within the nervous system predominantly in the cerebellum and the spinal cord while in peripheral tissues GFRalpha-3 was found to be expressed mostly in the colon, small intestine, pancreas, heart, testis and prostate. Using a GFRalpha-3-specific [35S]cRNA[gammaS] probe, in situ hybridization histochemistry experiments confirmed the expression in the cerebellum.
Masure
S
S
Department of Applied Molecular Biology, Janssen Research Foundation, Beerse, Belgium. smasure@janbe.jnj.com
Cik
M
M
Pangalos
M N
MN
Bonaventure
P
P
Verhasselt
P
P
Lesage
A S
AS
Leysen
J E
JE
Gordon
R D
RD
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D002874:Chromosome Mapping; D002895:Chromosomes, Human, Pair 5; D003001:Cloning, Molecular; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D007668:Kidney; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D009842:Oligopeptides; D009928:Organ Specificity; D010455:Peptides; D016133:Polymerase Chain Reaction; D051381:Rats; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection
C495039:GDNF protein, human; C494993:GFRA3 protein, human; C495041:Gdnf protein, rat; C494995:Gfra3 protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D009842:Oligopeptides; D010455:Peptides; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; C085706:FLAG peptide