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Originally published In Press as doi:10.1074/jbc.M107177200 on October 11, 2001

J. Biol. Chem., Vol. 276, Issue 52, 48978-48987, December 28, 2001
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Two Forms of Mitochondrial DNA Ligase III Are Produced in Xenopus laevis Oocytes*

Romina M. Perez-Jannotti, Seth M. Klein, and Daniel F. BogenhagenDagger

From the Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, New York 11794-8651

Full-length cDNAs for DNA ligase IV and the alpha  and beta  isoforms of DNA ligase III were cloned from Xenopus laevis to permit study of the genes encoding mitochondrial DNA ligase. DNA ligase IIIalpha and IIIbeta share a common NH2 terminus that encodes a mitochondrial localization signal capable of targeting green fluorescent protein to mitochondria while the NH2 terminus of DNA ligase IV does not. Reverse transcriptase-polymerase chain reaction analyses with adult frog tissues demonstrate that while DNA ligase IIIalpha and DNA ligase IV are ubiquitously expressed, DNA ligase IIIbeta expression is restricted to testis and ovary. Mitochondrial lysates from X. laevis oocytes contain both DNA ligase IIIalpha and IIIbeta but no detectable DNA ligase IV. Gel filtration, sedimentation, native gel electrophoresis, and in vitro cross-linking experiments demonstrate that mtDNA ligase IIIalpha exists as a high molecular weight complex. We discuss the possibility that DNA ligase IIIalpha exists in mitochondria in association with novel mitochondrial protein partners or as a homodimer.

* This work was supported by National Institutes of Health Grant RO1GM29681 and a W. Burghardt Turner Minority Fellowship Award (to R. M. P-J.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The cDNA sequences of X. laevis DNA ligase IIIalpha , DNA ligase IIIbeta , and DNA ligase IV have been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF393654, AF393655, and AF393656, respectively.

Dagger To whom correspondence should be addressed. Tel.: 631-444-3068; Fax: 631-444-3218; E-mail: dan@pharm.sunysb.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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