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Difference in Protein Expression in Vero Cells after Antisense-blocking Genes Involved in the Suppression of

J H Jin1, Y N Yu, Y L Qian

  • 1Department of Pathophysiology, Medical School of Zhejiang University, Hangzhou 310031, China. ynyu@mail.hz.zj.cn

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica Et Biophysica Sinica
|May 30, 2002
PubMed

Insights

The FNR gene may regulate cellular responses to DNA damage. Blocking FNR gene expression altered protein production in mammalian cells exposed to MNNG, suggesting its role in mutagenesis.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The fragment 9 related gene (FNR gene) is investigated for its potential role in inhibiting non-targeted mutagenesis.
  • N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a mutagen used to induce DNA damage in mammalian cells.

Purpose of the Study:

  • To elucidate the functional mechanism of the FNR gene by analyzing protein expression changes.
  • To understand the FNR gene's role in the cellular response to MNNG-induced mutagenesis.

Main Methods:

  • mRNA differential display and antisense technology were used for gene isolation.
  • Two-dimensional gel electrophoresis and 2D image analysis compared protein expression in transfected cells.
  • Vero cells were exposed to MNNG and transfected with antisense RNA expression plasmid or vector DNA.

Main Results:

  • Antisense blocking of FNR gene expression led to specific protein expression changes.
  • 12 proteins were uniquely expressed in cells with blocked FNR gene expression.
  • 24 proteins showed increased expression (P <0.05) in cells with blocked FNR gene expression, with 7 proteins enhanced >5-fold.

Conclusions:

  • FNR gene expression likely influences the expression of other genes.
  • The FNR gene may function as a regulatory factor in cellular responses to DNA damage.
  • This study aids in identifying proteins and genes involved in non-targeted mutagenesis.

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