Effects of RNA interference of Trypanosoma brucei structure-specific endonuclease-I on kinetoplast DNA replication.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Liu Y;Liu Y; Motyka SA; Englund PT
  • Source:
    The Journal of biological chemistry [J Biol Chem] 2005 Oct 21; Vol. 280 (42), pp. 35513-20. Date of Electronic Publication: 2005 Aug 11.
  • Publication Type:
    Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Print ISSN: 0021-9258 (Print) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE
    • Publication Information:
      Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
      Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
    • Subject Terms:
      RNA Interference*; DNA/*genetics ; DNA, Kinetoplast/*genetics ; Endonucleases/*physiology ; Trypanosoma brucei brucei/*genetics ; Trypanosoma brucei brucei/*metabolism; Animals ; Blotting, Northern ; DNA/chemistry ; DNA Primers/chemistry ; DNA Restriction Enzymes/metabolism ; DNA, Kinetoplast/metabolism ; Endonucleases/chemistry ; Gene Silencing ; Green Fluorescent Proteins/metabolism ; Microscopy, Fluorescence ; Protein Structure, Tertiary ; RNA/chemistry ; Time Factors
    • Abstract:
      Kinetoplast DNA, the mitochondrial DNA of trypanosomatid protozoa, is a network containing several thousand topologically interlocked DNA minicircles. Kinetoplast DNA synthesis involves release of minicircles from the network, replication of the free minicircles, and reattachment of the progeny back onto the network. One enzyme involved in this process is structure-specific endonuclease-I. This enzyme, originally purified from Crithidia fasciculata, has been proposed to remove minicircle replication primers (Engel, M. L., and Ray, D. S. (1998) Nucleic Acids Res. 26, 4773-4778). We have studied the structure-specific endonuclease-I homolog from Trypanosoma brucei, showing it to be localized in the antipodal sites flanking the kinetoplast DNA disk, as previously shown in C. fasciculata. RNA interference of structure-specific endonuclease-I caused persistence of a single ribonucleotide at the 5' end of both the leading strand and at least the first Okazaki fragment in network minicircles, demonstrating that this enzyme in fact functions in primer removal. Probably because of the persistence of primers, RNA interference also impeded the reattachment of newly replicated free minicircles to the network and caused a delay in kinetoplast DNA segregation. These effects ultimately led to shrinkage and loss of the kinetoplast DNA network and cessation of growth of the cell.
    • Grant Information:
      AI058613 United States AI NIAID NIH HHS
    • Accession Number:
      0 (DNA Primers)
      0 (DNA, Kinetoplast)
      147336-22-9 (Green Fluorescent Proteins)
      63231-63-0 (RNA)
      9007-49-2 (DNA)
      EC 3.1.- (Endonucleases)
      EC 3.1.21.- (DNA Restriction Enzymes)
      EC 3.1.21.- (structure-specific endonuclease I)
    • Publication Date:
      Date Created: 20050813 Date Completed: 20051213 Latest Revision: 20210209
    • Publication Date:
      20231215
    • Accession Number:
      10.1074/jbc.M507296200
    • Accession Number:
      16096280