Cloning, Sequence Analysis, and Expression of the Large Subunit of the Human Lymphocyte Activation Antigen 4F2

Among the earliest expressed antigens on the surface of activated human lymphocytes is the surface antigen 4F2. We have used DNA-mediated gene transfer and fluorescence-activated cell sorting to obtain cell lines that contain the gene encoding the large subunit of the human 4F2 antigen in a mouse L-...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-12, Vol.84 (24), p.9204-9208
Main Authors: Lumadue, Jeanne A., Glick, Adam B., Ruddle, Frank H.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Antigens
Antigens, Surface - genetics
B lymphocytes
Base Sequence
Biological and medical sciences
Biotechnology
Cell Cycle
Cell lines
Cloning, Molecular
Complementary DNA
DNA
DNA - genetics
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
Genetic engineering
Genetic technics
Humans
Lymphocyte Activation
Lymphocytes
Lymphocytes - physiology
Membrane Glycoproteins - genetics
Methods. Procedures. Technologies
Molecular cloning
Molecular Sequence Data
Proteins
RNA
T lymphocytes
Transfection
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Summary:Among the earliest expressed antigens on the surface of activated human lymphocytes is the surface antigen 4F2. We have used DNA-mediated gene transfer and fluorescence-activated cell sorting to obtain cell lines that contain the gene encoding the large subunit of the human 4F2 antigen in a mouse L-cell background. Human DNAs cloned from these cell lines were subsequently used as hybridization probes to isolate a full-length cDNA clone expressing 4F2. Sequence analysis of the coding region has revealed an amino acid sequence of 529 residues. Hydrophobicity plotting has predicted a probable structure for the protein that includes an external carboxyl terminus, an internal leader sequence, a single hydrophobic transmembrane domain, and two possible membrane-associated domains. The 4F2 cDNA detects a single 1.8-kilobase mRNA in T-cell and B-cell lines. RNA gel blot analysis of RNA derived from quiescent and serum-stimulated Swiss 3T3 fibroblasts reveals a cell-cycle modulation of 4F2 gene expression: the mRNA is present in quiescent fibroblasts but increases 8-fold 24-36 hr after stimulation, at the time of maximal DNA synthesis.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.24.9204