Mitochondrial DNA ligase in Crithidia fasciculata
- Molecular Biology Institute and Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095
-
Edited by Robert Lehman, Stanford University School of Medicine, Stanford, CA, and approved January 15, 2004 (received for review September 5, 2003)
Abstract
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).
Footnotes
-
↵ * To whom correspondence should be addressed at: 301A Paul D. Boyer Hall, 611 Charles Young Drive East, University of California, Los Angeles, CA 90095-1570. E-mail: danray{at}ucla.edu.
-
This paper was submitted directly (Track II) to the PNAS office.
-
Abbreviations: HA, hemagglutinin; kDNA, kinetoplast DNA.
-
Data deposition: The sequence reported in this paper has been deposited in the GenBank database (accession no. AY380335).
-
See Commentary on page 4333.
- Copyright © 2004, The National Academy of Sciences
