cDNA and protein characterization of human MAGE-10.
MAGE genes are frequently expressed in several types of human malignancy and code for antigens recognized by cytotoxic T lymphocytes. We have previously described a monoclonal antibody (MAb), named 6C1, that recognizes the MAGE-1 protein and cross-reacts with a 72-kDa protein present in lysates of melanoma cells such as MZ2-MEL. To identify this protein, we have screened an expression library prepared from MZ2-MEL cells. Several clones that encoded a protein recognized by antibody 6C1 contained a sequence identical to that of MAGE-10, another member of the MAGE-A gene family. Full-length MAGE-10 cDNA clones, obtained after screening additional cDNA melanoma libraries, were found to be approximately 2.5 kb in length. In vitro translation and transient transfection experiments indicated that MAGE-10 codes for a protein of approximately 72 kDa. This product was recognized by MAb 6C1 as well as by a polyclonal serum raised against a MAGE-10 peptide, thus demonstrating its identity with MAGE-10. Analysis of MAGE-10 mRNA by RT-PCR confirmed its presence in testis and placenta but not in other normal tissues. Expression of MAGE-10 in melanoma tumors was found to parallel that of MAGE-1. Western blot analysis with the polyclonal anti-MAGE-10 antibody showed the presence of MAGE-10 in lysates of purified trophoblast cells. Immuno-cytochemistry of cultured melanoma cells indicated that MAGE-10 is a nuclear protein.
Rimoldi
D
D
Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges, Switzerland. drimoldi@eliot.unil.ch
Salvi
S
S
Reed
D
D
Coulie
P
P
Jongeneel
V C
VC
De Plaen
E
E
Brasseur
F
F
Rodriguez
A M
AM
Boon
T
T
Cerottini
J C
JC
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000235:genetics*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D015723:Gene Library; D006801:Humans; D007438:Introns; D008297:Male; D008545:Melanoma / Q000235:genetics*; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D018076:DNA, Complementary; C121012:MAGEA10 protein, human; D009363:Neoplasm Proteins; D011994:Recombinant Proteins