Kinetoplast-associated DNA topoisomerase in Crithidia fasciculata: crosslinking of mitochondrial topoisomerase II to both minicircles and maxicircles in cells treated with the topoisomerase inhibitor VP16.
The mitochondrial DNA of the trypanosomatid Crithidia fasciculata consists of thousands of copies of a 2.5 kb minicircle and a small number of 37kb maxicircles catenated into a single enormous network. Treatment of C. fasciculata with the type II DNA topoisomerase inhibitor VP16 produces cleavable complexes of a type II DNA topiosomerase with both minicircles and maxicircles. A combined Southern and Western blot analysis of the cleaved DNA species released from the network by SDS treatment has identified topollmt, the kinetoplast-associated topisomerase, in covalent complexes with linear forms of minicircle and maxicircle DNAs. These results directly implicate topollmt in the topological reactions required for the duplication of the kinetoplast network.
Ray
D S
DS
Molecular Biology Institute, UCLA 90024.
Hines
J C
JC
Anderson
M
M
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D016831:Crithidia fasciculata / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004270:DNA, Circular / Q000378:metabolism*; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan / Q000378:metabolism*; D005047:Etoposide / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology*
D004270:DNA, Circular; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D005047:Etoposide; D004250:DNA Topoisomerases, Type II