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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
07:09

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

Published on: July 31, 2012

De novo protein synthesis is essential to human interferon gamma gene expression by the stimulation with polyI:polyC

M Tamura-Nishimura1, S Sasakawa

  • 1Japanese Red Cross, Central Blood Center, Tokyo.

FEBS Letters
|February 26, 1990
PubMed

Insights

De novo protein synthesis is essential for early interferon (IFN) gamma gene expression. Inhibiting protein synthesis prevents IFN-gamma mRNA transcription, suggesting a crucial role for newly synthesized proteins in gene induction.

Area of Science:

  • Immunology
  • Molecular Biology
  • Gene Expression

Background:

  • Human interferon (IFN) gamma gene expression is induced by double-stranded RNA (poly I:poly C) in peripheral lymphocyte NNA cells.
  • Early gene expression requires de novo protein synthesis.

Purpose of the Study:

  • To investigate the necessity of de novo protein synthesis in the early stages of IFN-gamma gene expression.
  • To identify the role of protein synthesis inhibitors in IFN-gamma induction.

Main Methods:

  • Treatment of NNA cells with poly I:poly C and cycloheximide (CHX), a protein synthesis inhibitor.
  • Cell-free translation assays using RNA isolated from treated cells.

Main Results:

  • A 4-hour treatment with poly I:poly C is sufficient for IFN-gamma gene induction.
  • Addition of CHX during the initial 4 hours of induction inhibits IFN-gamma induction.
  • IFN-gamma mRNA was absent in cells treated with poly I:poly C and CHX, indicating blocked transcription.

Conclusions:

  • De novo protein synthesis is required for the transcription of IFN-gamma mRNA.
  • The essential protein(s) may be involved in protein kinase C (pkC) activation, which is crucial for IFN-gamma gene induction.

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