Abstract
IEX-1L has been claimed to act as an apoptosis inhibitor involved in NFkappaB-mediated survival in Jurkat cells [Wu et al. (1998) Science 281, 998-1001]. It represents a mutant nonspliced variant of the early response gene p22(PRG1/IEX-1) exhibiting one insertion and two deletions compared to the genomic sequence of p22(PRG1/IEX-1). Direct DNA sequencing of PCR products generated from human genomic DNA only detected the regular genomic sequence of p22(PRG1/IEX-1). No IEX-1L mRNA could be identified by RT-PCR analysis and subsequent DNA sequencing of total, nuclear, or cytoplasmic RNA fractions from PMA-stimulated Jurkat cells. The only functional transcript residing in the cytoplasm is regularly spliced p22(IEX-1/PRG1) mRNA. Substantial amounts of nonmutated nonspliced p22(IEX-1/PRG1) pre-mRNA were identified in the nucleus. Thus, IEX-1L seems to be a mutant variant of p22(IEX-1/PRG1) not existing in vivo. Antiapoptotic effects obviously represent transdominant negative inhibition of endogenous p22(PRG1/IEX-1) in Jurkat cells and several other tumor cell lines.
Copyright 1999 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alternative Splicing / drug effects
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Alternative Splicing / genetics*
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Animals
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Apoptosis Regulatory Proteins
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Apoptosis*
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Base Sequence
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Cell Line
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Cell Nucleus / drug effects
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Cell Nucleus / genetics
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Cell Survival
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Cricetinae
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Cytoplasm / drug effects
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Cytoplasm / genetics
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Gene Expression* / drug effects
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Humans
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Immediate-Early Proteins / chemistry
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Immediate-Early Proteins / genetics*
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Immediate-Early Proteins / metabolism*
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Membrane Glycoproteins / chemistry
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Membrane Glycoproteins / genetics*
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Membrane Glycoproteins / metabolism*
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Membrane Proteins
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Molecular Sequence Data
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Mutation*
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NF-kappa B / metabolism
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Neoplasm Proteins*
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Protein Isoforms / chemistry
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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RNA, Messenger / analysis
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RNA, Messenger / genetics
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Tetradecanoylphorbol Acetate / pharmacology
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Transfection
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Tumor Necrosis Factor-alpha / pharmacology
Substances
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Apoptosis Regulatory Proteins
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IER3 protein, human
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Immediate-Early Proteins
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Membrane Glycoproteins
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Membrane Proteins
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NF-kappa B
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Neoplasm Proteins
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Protein Isoforms
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RNA, Messenger
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Tumor Necrosis Factor-alpha
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Tetradecanoylphorbol Acetate