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Mol. Cell. Biol., 10 1995, 5412-5422, Vol 15, No. 10
J Chen, AE Tomkinson, W Ramos, ZB Mackey, S Danehower, CA Walter, RA Schultz, JM Besterman and I Husain
Three biochemically distinct DNA ligase activities have been identified in
mammalian cell extracts. We have recently purified DNA ligase II and DNA
ligase III to near homogeneity from bovine liver and testis tissue,
respectively. Amino acid sequencing studies indicated that these enzymes
are encoded by the same gene. In the present study, human and murine cDNA
clones encoding DNA ligase III were isolated with probes based on the
peptide sequences. The human DNA ligase III cDNA encodes a polypeptide of
862 amino acids, whose sequence is more closely related to those of the DNA
ligases encoded by poxviruses than to replicative DNA ligases, such as
human DNA ligase I. In vitro transcription and translation of the cDNA
produced a catalytically active DNA ligase similar in size and substrate
specificity to the purified bovine enzyme. The DNA ligase III gene was
localized to human chromosome 17, which eliminated this gene as a candidate
for the cancer-prone disease Bloom syndrome that is associated with DNA
joining abnormalities. DNA ligase III is ubiquitously expressed at low
levels, except in the testes, in which the steady-state levels of DNA
ligase III mRNA are at least 10-fold higher than those detected in other
tissues and cells. Since DNA ligase I mRNA is also present at high levels
in the testes, we examined the expression of the DNA ligase genes during
spermatogenesis. DNA ligase I mRNA expression correlated with the
contribution of proliferating spermatogonia cells to the testes, in
agreement with the previously defined role of this enzyme in DNA
replication. In contrast, elevated levels of DNA ligase III mRNA were
observed in primary spermatocytes undergoing recombination prior to the
first meiotic division. Therefore, we suggest that DNA ligase III seals DNA
strand breaks that arise during the process of meiotic recombination in
germ cells and as a consequence of DNA damage in somatic cells.
Copyright © 1995, American Society for Microbiology
Mammalian DNA ligase III: molecular cloning, chromosomal localization, and expression in spermatocytes undergoing meiotic recombination
Department of Cell Biology, Glaxo Research Institute, Research Triangle Park, North Carolina 27709, USA.
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