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Alternative RNA splicing in expression of the glutathione synthetase gene in human cells

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Abstract

Ubiquitous free radical production occurs continuously in cells and tissues. Glutathione is the most abundant mammalian antioxidant, and is synthesized by glutathione synthetase (GSS). Therefore, GSS plays an important role in defending the cell against reactive oxygen species. The expression of GSS has been studied in human cells; however, sequence information about alternative splicing variants of GSS mRNA has not been reported. In the present study, we identified a novel alternative splicing variant (ASV) of the GSS gene in 10 human normal tissues and five human cancer cell lines. The deleted transcript of GSS was characterized by an in-frame deletion of 333 bp, corresponding to the complete loss of exons 4 and 5. Thus this GSS ASV causes protein truncation. We quantified the mRNA of GSS ASV in human normal tissues using real-time PCR. The ASV was detected in colon, kidney, lung, liver, placenta, peripheral blood and uterus, but not in heart, skeletal muscle and spleen tissue. Our results provide a basis for more detailed studies on the regulation of GSS, and for further evaluation of this and other possible roles of GSS. Understanding the regulation of GSS expression is very important for the development of new strategies for controlling the development of GSH-based redox homeostasis.

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Abbreviations

GSS:

Glutathione synthetase

GSH:

Glutathione

ASV:

Alternative splicing variant

cDNA:

Complementary DNA

RACE:

Rapid amplification of cDNA ends

ROS:

Reactive oxygen species

NMD:

Nonsense-mediated mRNA decay

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Acknowledgments

We are indebted to Naoko Shirakawa and Nanae Ishikawa of the 366 Rehabilitation Hospital for their kind assistance in this study.

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Authors and Affiliations

  1. Laboratory of Cellular and Molecular Biochemistry, Department of Materials and Life Science, Seikei University, 3-3-1 Kichijoji Kita-machi, Musashino-shi, Tokyo, 180-8633, Japan

    Marie Uchida, Maki Sugaya & Hisashi Hisatomi

  2. Department of Surgery, Kobe Rousai Hospital, Kobe, 651-0053, Japan

    Taichi Kanamaru

Authors
  1. Marie Uchida
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  2. Maki Sugaya
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  3. Taichi Kanamaru
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  4. Hisashi Hisatomi
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Corresponding author

Correspondence to Hisashi Hisatomi.

Additional information

The sequence reported in this paper has been deposited in the GenBank database (accession no. AB459500).

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Uchida, M., Sugaya, M., Kanamaru, T. et al. Alternative RNA splicing in expression of the glutathione synthetase gene in human cells. Mol Biol Rep 37, 2105–2109 (2010). https://doi.org/10.1007/s11033-009-9675-3

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  • DOI: https://doi.org/10.1007/s11033-009-9675-3

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