Biomedical Science Letters (대한의생명과학회지)

The Korean Society for Biomedical Laboratory Sciences (대한의생명과학회)
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Evidence of DNA Replication Licensing and Paternal DNA Degradation by MCM7 and ORC2 in the Mouse One-cell Embryo

  • Kim, Chang Jin (Department of Biology Education, College of Education, Chonnam National University) ;
  • Kim, Tae Hoon (Department of Food Science and Biotechnology, College of Engineering, Daegu University) ;
  • Lee, Eun-Woo (Division of Applied Bioengineering, College of Engineering, Dong-eui University) ;
  • Lee, Kyung-Bon (Department of Biology Education, College of Education, Chonnam National University)
  • Received : 2017.10.13
  • Accepted : 2017.11.07
  • Published : 2017.12.31
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Abstract

This study was investigated to test whether paternal DNA that was destined for degradation was properly licensed by testing for the presence of mini-chromosome maintenance protein (MCM) 7 and origin recognition complex (ORC) 2 in the paternal pronuclei. ORC2 is one of the first licensing protein to come on and MCM7 is one of the last licensing protein to come on. Zygotes were prepared by injection of control and treated sperm injection (ICSI). To control for DNA breakage, epididymal spermatozoa were treated with DNase I to fragment the DNA, then injected into oocytes. The presence of MCM7 and ORC2 in the pronuclei of mouse zygotes was tested by immunohistochemistry, just before the onset of DNA synthesis, at 5 h after fertilization, and after DNA synthesis began, at 9 h post fertilization. We found that in all cases, both MCM7 and ORC2 were present in both pronuclei at 5 h after sperm injection, just before DNA synthesis began. This indicates that no matter how extensive the DNA damage, recruitment of licensing proteins to the origins of replication was not inhibited. Sperm DNA fragmentation does not prevent licensing of DNA replication origins. Furthermore, the embryo recognizes DNA that is damaged by nucleases. Our data indicate that the one-cell embryo does harbor a mechanism to prevent the replication of severely damaged DNA from spermatozoa, even though the embryos do not undergo classical apoptosis.

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References

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