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PMID: 2153072 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Expression and targeting of intracellular antibodies in mammalian cells.

The EMBO journal ·Vol. 9 ·No. 1 ·1990-01-00 ·Pages 101-8

Biocca S, Neuberger MS, Cattaneo A

Abstract

Genes encoding the heavy and light chains of a hapten-specific IgM antibody were modified by site-directed mutagenesis to destroy the hydrophobic leader sequences and allow expression in the cytoplasm of non-lymphoid cells. The in situ assembly of the mutant heavy and light chains was tested in transfected cell lines by immunofluorescence using anti-idiotypic antibodies. A positive diffuse cytoplasmic staining was observed. This demonstrated that the antibody polypeptide chains could assemble in the cell cytoplasm and led us to ask whether antibodies could be further targeted to the nucleus. Mutations were therefore made in which the leader sequence of the light chain was replaced by the nuclear localization signal of the SV40 large T antigen. Transfectants in which the heavy chain lacking the hydrophobic leader was expressed together with a light chain carrying the nuclear localization signal were selected and a nuclear distribution of the assembled antibody was found. Thus, it should prove possible to target a specific antibody to the cell nucleus with the aim of interfering with the function of a nuclear antigen.

MeSH Terms
Amino Acid Sequence Animals Antigens, Viral, Tumor/genetics Base Sequence Cell Nucleus/immunology Cytoplasm/immunology Fluorescent Antibody Technique Gene Expression Genes, Immunoglobulin Haptens Immunoglobulin Heavy Chains/genetics Immunoglobulin Light Chains/genetics Immunoglobulin M/genetics Mice Molecular Sequence Data Mutation Plasmids Protein Sorting Signals/genetics Rats Simian virus 40/immunology Transfection Tumor Cells, Cultured
Chemicals
Antigens, Viral, Tumor Haptens Immunoglobulin Heavy Chains Immunoglobulin Light Chains Immunoglobulin M Protein Sorting Signals
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Biocca S
Institute of Neurobiology, CNR, Roma, Italy.
Neuberger M S
Cattaneo A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-01-00
Pages
101-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551635
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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