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Molecular and Cellular Biology, July 2006, p. 5382-5393, Vol. 26, No. 14
0270-7306/06/$08.00+0     doi:10.1128/MCB.00369-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Role of p38 in Replication of Trypanosoma brucei Kinetoplast DNA

Beiyu Liu,1 Henrik Molina,1 Dario Kalume,1 Akhilesh Pandey,1 Jack D. Griffith,2 and Paul T. Englund1*

Department of Biological Chemistry, Johns Hopkins Medical School, Baltimore, Maryland 21205,1 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 275992

Received 1 March 2006/ Returned for modification 29 March 2006/ Accepted 5 May 2006

Trypanosomes have an unusual mitochondrial genome, called kinetoplast DNA, that is a giant network containing thousands of interlocked minicircles. During kinetoplast DNA synthesis, minicircles are released from the network for replication as {theta}-structures, and then the free minicircle progeny reattach to the network. We report that a mitochondrial protein, which we term p38, functions in kinetoplast DNA replication. RNA interference (RNAi) of p38 resulted in loss of kinetoplast DNA and accumulation of a novel free minicircle species named fraction S. Fraction S minicircles are so underwound that on isolation they become highly negatively supertwisted and develop a region of Z-DNA. p38 binds to minicircle sequences within the replication origin. We conclude that cells with RNAi-induced loss of p38 cannot initiate minicircle replication, although they can extensively unwind free minicircles.

* Corresponding author. Mailing address: Department of Biological Chemistry, Johns Hopkins Medical School, 725 N. Wolfe St., Baltimore, MD 21205. Phone: (410) 955-3790. Fax: (410) 955-7810. E-mail: penglund{at}jhmi.edu.

Molecular and Cellular Biology, July 2006, p. 5382-5393, Vol. 26, No. 14
0270-7306/06/$08.00+0     doi:10.1128/MCB.00369-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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