Subcellullar localization of tumor-associated antigen 3H11Ag.
3H11Ag, a tumor-associated antigen defined by the monoclonal antibody 3H11 that specifically recognizes cancer cells in various tumor tissues, was successfully cloned recently, but its function is unknown. To explore the potential roles it plays in tumors, we analyzed its subcellular localization in the present study. By expressing 3H11Ag fused with fluorescent protein in COS-7 cells, we found that 3H11Ag localizes to both cytoplasm and nucleus, which was confirmed by subcellular fractionation. And sequentially extracting the nuclei of COS-7 cells transfected with 3H11Ag showed that it is a DNA- and nuclear matrix-associated protein. Moreover, by expressing a series of red fluorescent protein-tagged truncated forms of 3H11Ag, it was demonstrated that the 150 amino acid residues at its C-terminal are fully responsible for the subcellular localization. In addition, the results of the computational analysis of 3H11Ag were in accordance with those of the experimental analysis. All these data would be helpful to elucidate the functions of 3H11Ag.
Department of Biochemistry and Molecular Biology, Beijing Institute for Cancer Research, Peking University School of Oncology, Beijing, PR China.
Jianhui
J
Guo
Genglin
G
Jin
Lin
L
Meng
Hong
H
Ma
Dezhi
D
Nie
Jian
J
Wu
Lan
L
Yuan
Chengchao
C
Shou
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000737:chemistry*; D015153:Blotting, Western; D019556:COS Cells; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000378:metabolism; D019295:Computational Biology; D003593:Cytoplasm / Q000378:metabolism; D003598:Cytoskeletal Proteins; D004247:DNA / Q000737:chemistry; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012984:Software; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D000906:Antibodies; D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C493903:Cep290 protein, human; D003598:Cytoskeletal Proteins; D009363:Neoplasm Proteins; D004247:DNA; D005982:Glutathione Transferase