Apoptosis

, Volume 3, Issue 2, pp 89–95

cDNA Cloning of Human DNase γ: Chromosomal Localization of Its Gene and Enzymatic Properties of Recombinant Protein

  • D. Shiokawa
  • M. Hirai
  • S. Tanuma
Article

DOI: 10.1023/A:1009692807692

Cite this article as:
Shiokawa, D., Hirai, M. & Tanuma, S. Apoptosis (1998) 3: 89. doi:10.1023/A:1009692807692

Abstract

We report here on the nucleotide sequence of the cDNA encoding human DNase γ, which is a candidate for an apoptotic endonuclease. The cDNA clone isolated from a human spleen cDNA library is composed of a 918 bp open reading frame encoding a 305 amino acid precursor protein for DNase γ. Northern blot analysis reveals that the expression of a single transcript of 1.5 kb DNase γ mRNA is detected in the spleen and liver. The chromosomal localization of DNase γ gene is mapped to chromosome 3 at region p21.1-p14.2 by fluorescence in situ hybridization (FISH). Characterization of thioredoxin-DNase γ fusion protein (Trx-hDNase γ) shows that the recombinant protein has a Ca2+/Mg2+- or Mn2+-dependent endonuclease activity that cleaves chromatin DNA to nucleosomal units. The optimum pH is around 7.2. Zn2+ and aurintricarboxylic acid (ATA) inhibits the activity in dose-dependent manners. These properties are identical to those of purified DNase γ.

ApoptosisDNA fragmentationDNase γendonuclease

Copyright information

© Rapid Science Ltd 1998

Authors and Affiliations

  • D. Shiokawa
    • 1
  • M. Hirai
    • 2
  • S. Tanuma
    • 1
  1. 1.Department of Biochemistry, Faculty of Pharmaceutical SciencesScience University of TokyoShinjuku-ku, TokyoJapan
  2. 2.Department of Biological Science, Graduate School of ScienceUniversity of TokyoBunkyo-ku, TokyoJapan