Identification and expression of two forms of the human transforming growth factor-beta-binding protein endoglin with distinct cytoplasmic regions.
Endoglin is an homodimeric membrane antigen with capacity to bind transforming growth factor-beta (TGF-beta) and whose expression is up-regulated on myeloid cells upon differentiation to macrophages. We have isolated full-length cDNA clones from a lambda gt 10 library, prepared from phorbol 12-myristate 13-acetate-differentiated HL60 cells by screening with an endoglin-specific cDNA probe from endothelial cells. Sequencing of the largest clone (3073 bp), revealed that the leader sequence contains 25 residues and that the 586 amino acids of the extracellular and transmembrane domains were identical to those described for endothelial endoglin. However, the cytoplasmic tail encoded by this cDNA clone contains only 14 amino acids as opposed to the 47 residues previously reported, suggesting the existence of two alternative endoglin variants. The expression of these isoforms was demonstrated by polymerase chain reaction analyses on endothelial cells, myelomonocytic cell lines HL-60 and U-937, and placenta. Independent cDNA constructs corresponding to both forms were transfected into mouse fibroblasts leading to the expression of two distinct endoglin molecules. Both forms were shown to bind TGF-beta 1 and, when overexpressed in transfected mouse fibroblasts, to form disulfide-linked homodimers, indicating that the cysteine residues present in the extracellular domain are responsible for the dimerization.
Bellón
T
T
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.
Corbí
A
A
Lastres
P
P
Calés
C
C
Cebrián
M
M
Vera
S
S
Cheifetz
S
S
Massague
J
J
Letarte
M
M
Bernabéu
C
C
D000595:Amino Acid Sequence; D015703:Antigens, CD; D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis*; D003593:Cytoplasm / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification; D000071063:Endoglin; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D053779:Latent TGF-beta Binding Proteins; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011956:Receptors, Cell Surface; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D019010:Vascular Cell Adhesion Molecule-1
D015703:Antigens, CD; D002352:Carrier Proteins; C064206:ENG protein, human; D000071063:Endoglin; D047908:Intracellular Signaling Peptides and Proteins; D053779:Latent TGF-beta Binding Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D016212:Transforming Growth Factor beta; D019010:Vascular Cell Adhesion Molecule-1; D004247:DNA