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Small dna pieces in C. elegans are intermediates of DNA fragmentation during apoptosis

Aruscavage, PJ and Hellwig, S and Bass, BL (2010) Small dna pieces in C. elegans are intermediates of DNA fragmentation during apoptosis. PLoS ONE, 5 (6).

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While studying small noncoding RNA in C. elegans, we discovered that protocols used for isolation of RNA are contaminated with small DNA pieces. After electrophoresis on a denaturing gel, the DNA fragments appear as a ladder of bands, 10 nucleotides apart, mimicking the pattern of nuclease digestion of DNA wrapped around a nucleosome. Here we show that the small DNA pieces are products of the DNA fragmentation that occurs during apoptosis, and correspondingly, are absent in mutant strains incapable of apoptosis. In contrast, the small DNA pieces are present in strains defective for the engulfment process of apoptosis, suggesting they are produced in the dying cell prior to engulfment. While the small DNA pieces are also present in a number of strains with mutations in predicted nucleases, they are undetectable in strains containing mutations in nuc-1, which encodes a DNase II endonuclease. We find that the small DNA pieces can be labeled with terminal deoxynucleotidyltransferase only after phosphatase treatment, as expected if they are products of DNase II cleavage, which generates a 39 phosphate. Our studies reveal a previously unknown intermediate in the process of apoptotic DNA fragmentation and thus bring us closer to defining this important pathway.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Aruscavage, PJ
Hellwig, Ssah89@pitt.eduSAH89
Bass, BL
ContributionContributors NameEmailPitt UsernameORCID
Date: 11 August 2010
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 5
Number: 6
DOI or Unique Handle: 10.1371/journal.pone.0011217
Refereed: Yes
MeSH Headings: 3' Untranslated Regions; Animals; Apoptosis; Caenorhabditis elegans--cytology; Caenorhabditis elegans--genetics; DNA Nucleotidylexotransferase--metabolism; DNA, Helminth--genetics
Other ID: NLM PMC2887891
PubMed Central ID: PMC2887891
PubMed ID: 20585459
Date Deposited: 03 Aug 2012 21:00
Last Modified: 04 Feb 2019 19:55


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