Resolution of recombination intermediates by a mammalian activity functionally analogous to Escherichia coli RuvC resolvase.
A mammalian endonuclease that resolves Holliday junctions has been partially purified from extracts of calf thymus and Chinese hamster ovary cells. The activity acts upon (i) synthetic Holliday junctions and (ii) recombination intermediates made by the Escherichia coli RecA protein and appears to be functionally analogous to the E. coli RuvC protein. Cleavage occurs by the introduction of symmetrically related nicks in strands of like polarity to produce nicked duplex DNA products. The nicks can be repaired by DNA ligase. The resolvase is specific for Holliday junctions and does not act upon Y junctions, G/A mismatches, or heterologous loops. The substrate specificity is therefore similar to that of E. coli RuvC protein and contrasts with the broad range specificity of other junction resolvases such as T4 endonuclease VII. The mammalian resolvase activity has been observed at normal levels in extracts prepared from a series of DNA repair-defective cells. These include the x-ray or UV-sensitive hamster lines xrs-5, xrs-6, and Chinese hamster ovary 43-3B (defective in ERCC-1), and murine cells that are severely immunodeficient and defective in both V(D)J rejoining and DNA repair.
Hyde
H
H
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire, United Kingdom.
Davies
A A
AA
Benson
F E
FE
West
S C
SC
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D016466:CHO Cells; D002417:Cattle; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D004279:DNA, Viral / Q000302:isolation & purification / Q000378:metabolism; D004706:Endodeoxyribonucleases; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D011995:Recombination, Genetic; D013379:Substrate Specificity; D013950:Thymus Gland / Q000378:metabolism*; D019895:Transposases
D001426:Bacterial Proteins; D004279:DNA, Viral; D029968:Escherichia coli Proteins; D009838:Oligodeoxyribonucleotides; C069806:ruvC protein, E coli; D009713:Nucleotidyltransferases; D019895:Transposases; D004706:Endodeoxyribonucleases