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J. Biol. Chem., Vol. 281, Issue 49, 37468-37476, December 8, 2006

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Binding of the Universal Minicircle Sequence Binding Protein at the Kinetoplast DNA Replication Origin^*

Itay Onn¹, Irit Kapeller, Kawther Abu-Elneel², and Joseph Shlomai³

From the Department of Parasitology, The Kuvin Center for the Study of Infectious and Tropical Diseases and the Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel

Kinetoplast DNA, the mitochondrial DNA of trypanosomatids, is a remarkable DNA structure that contains, in the species Crithidia fasciculata, 5000 topologically linked duplex DNA minicircles. Their replication initiates at two conserved sequences, a dodecamer, known as the universal minicircle sequence (UMS), and a hexamer, which are located at the replication origins of the minicircle L and H strands, respectively. A UMS-binding protein (UMSBP) binds specifically the 12-mer UMS sequence and a 14-mer sequence that contains the conserved hexamer in their single-stranded DNA conformation. In vivo cross-linking analyses reveal the binding of UMSBP to kinetoplast DNA networks in the cell. Furthermore, UMSBP binds in vitro to native minicircle origin fragments, carrying the UMSBP recognition sequences. UMSBP binding at the replication origin induces conformational changes in the bound DNA through its folding, aggregation and condensation.

Received for publication, July 5, 2006 , and in revised form, September 19, 2006.

^* This study was supported in part by United State-Israel Binational Science Foundation (Jerusalem, Israel) Grant 2001006 and by Israel Science Foundation Grant 623. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Tables 1 and 2.

¹ Supported by a Yeshaya Horovitz Fellowship. Current address: Cold Spring Harbor Laboratory, 1 Bungtown Rd., Cold Spring Harbor, NY 11724.

² Supported by a fellowship from the Ministry of Science, Israel. Current address: University of California Santa Barbara, Neuroscience Research Institute, Santa Barbara, CA 93106.

³ To whom correspondence should be addressed. Tel.: 972-2-6758089; Fax: 972-2-6757425; E-mail: shlomai{at}cc.huji.ac.il.

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