Purification and characterization of DNA ligase III from bovine testes. Homology with DNA ligase II and vaccinia DNA ligase.
Mammalian cell nuclei contain three biochemically distinct DNA ligases. In the present study we have found high levels of DNA ligase I and DNA ligase III activity in bovine testes and have purified DNA ligase III to near homogeneity. The high level of DNA ligase III suggests a role for this enzyme in meiotic recombination. In assays measuring the fidelity of DNA joining, we detected no significant differences between DNA ligases II and III, whereas DNA ligase I was clearly a more faithful enzyme and was particularly sensitive to 3' mismatches. Amino acid sequences of peptides derived from DNA ligase III demonstrated that this enzyme, like DNA ligase II, is highly homologous with vaccinia DNA ligase. The absence of unambiguous differences between homologous peptides from DNA ligases II and III (10 pairs of peptides, 136 identical amino acids) indicates that these enzymes are either derived from a common precursor polypeptide or are encoded from the same gene by alternative splicing. Based on similarities in amino acid sequence and biochemical properties, we suggest that DNA ligases II and III, Drosophila DNA ligase II, and the DNA ligases encoded by the pox viruses constitute a distinct family of DNA ligases that perform specific roles in DNA repair and genetic recombination.
Husain
I
I
Department of Cell Biology, Glaxo Research Institute, Research Triangle Park, North Carolina 27709, USA.
Tomkinson
A E
AE
Burkhart
W A
WA
Moyer
M B
MB
Ramos
W
W
Mackey
Z B
ZB
Besterman
J M
JM
Chen
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002467:Cell Nucleus / Q000201:enzymology; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D007106:Immune Sera / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D000075223:Poly-ADP-Ribose Binding Proteins; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000201:enzymology*; D014616:Vaccinia virus / Q000201:enzymology*; D029867:Xenopus Proteins
D007106:Immune Sera; D000075223:Poly-ADP-Ribose Binding Proteins; D029867:Xenopus Proteins; D011088:DNA Ligases; D000072481:DNA Ligase ATP; C000618816:DNA ligase III alpha protein, Xenopus