Journal
MOLECULAR CELL
Volume 35, Issue 4, Pages 490-501Publisher
CELL PRESS
DOI: 10.1016/j.molcel.2009.07.004
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Funding
- NIH [AI058613, 2005023]
- United States-Israel Binational Science Foundation [54/06]
- Israel Science Foundation
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Kinetoplast DNA (kDNA), the trypanosome mitochondrial DNA, contains thousands of minicircles and dozens of maxicircles interlocked in a giant network. Remarkably, Trypanosoma brucei's genome encodes 8PIF1-like helicases, 6 of which are mitochondrial. We now show that TbPIF2 is essential for maxicircle replication. Maxicircle abundance is controlled by TbPIF2 level, as RNAi of this helicase caused maxicircle loss, and its overexpression caused a 3- to 6-fold increase in maxicircle abundance. This regulation of maxicircle level is mediated by the TbHsIVU protease. Previous experiments demonstrated that RNAi knockdown of TbHsIVU dramatically increased abundance of minicircles and maxicircles, presumably because a positive regulator of their synthesis escaped proteolysis and allowed synthesis to continue. Here, we found that TbPIF2 level increases following RNAi of the protease. Therefore, this helicase is a TbHsIVU substrate and an example of a positive regulator, thus providing a molecular mechanism for controlling maxicircle replication.
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