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Mitochondrial DNA ligase in Crithidia fasciculata

Kinetoplast DNA (kDNA), the form of mitochondrial DNA in trypanosomatids, consists of thousands of interlocked circular DNAs organized into a compact disk structure. A type II DNA topoisomerase, a DNA polymerase β, and a structure-specific endonuclease have been localized to antipodal sites flanking...

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Detalhes bibliográficos
Main Authors: Sinha, Krishna Murari, Hines, Jane C., Downey, Nicholas, Ray, Dan S.
Formato: Artigo
Idioma:en
Publicado em: National Academy of Sciences 2004
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Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC384752/
https://ncbi.nlm.nih.gov/pubmed/15070723
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1073/pnas.0305705101
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Resumo:Kinetoplast DNA (kDNA), the form of mitochondrial DNA in trypanosomatids, consists of thousands of interlocked circular DNAs organized into a compact disk structure. A type II DNA topoisomerase, a DNA polymerase β, and a structure-specific endonuclease have been localized to antipodal sites flanking the kDNA disk along with nascent DNA minicircles. We have cloned a gene (LIG k) encoding a mitochondrial DNA ligase in the trypanosomatid Crithidia fasciculata, and we show that an epitope-tagged form of the ligase colocalizes with the other replication proteins at the antipodal sites and also at the two faces of the kDNA disk. DNA LIG k becomes adenylated in reactions with ATP, and the adenylate moiety is removed by incubation with pyrophosphate or nicked DNA. The ligase interacts physically with the β polymerase and is proposed to be involved in the repair of gaps in the newly synthesized minicircles. In yeast and mammals, a single gene encodes both nuclear and mitochondrial forms of DNA ligase. The LIG K protein sequence has low similarity to mitochondrial DNA ligases in other eukaryotes and is distinct from the C. fasciculata nuclear DNA ligase (LIG I).