Leishmania actin binds and nicks kDNA as well as inhibits decatenation activity of type II topoisomerase.
Leishmania actin (LdACT) is an unconventional form of eukaryotic actin in that it markedly differs from other actins in terms of its filament forming as well as toxin and DNase-1-binding properties. Besides being present in the cytoplasm, cortical regions, flagellum and nucleus, it is also present in the kinetoplast where it appears to associate with the kinetoplast DNA (kDNA). However, nothing is known about its role in this organelle. Here, we show that LdACT is indeed associated with the kDNA disc in Leishmania kinetoplast, and under in vitro conditions, it specifically binds DNA primarily through electrostatic interactions involving its unique DNase-1-binding region and the DNA major groove. We further reveal that this protein exhibits DNA-nicking activity which requires its polymeric state as well as ATP hydrolysis and through this activity it converts catenated kDNA minicircles into open form. In addition, we show that LdACT specifically binds bacterial type II topoisomerase and inhibits its decatenation activity. Together, these results strongly indicate that LdACT could play a critical role in kDNA remodeling.
Division of Molecular and Structural Biology, Central Drug Research Institute, Chattar Manzil Palace, CSIR, Lucknow 226001, India.
Prabodh
P
Kapoor
Ashutosh
A
Kumar
Rangeetha
R
Naik
Munia
M
Ganguli
Mohammad I
MI
Siddiqi
Amogh A
AA
Sahasrabuddhe
Chhitar M
CM
Gupta
D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002843:Chromatin; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D043864:DNA, Catenated / Q000378:metabolism; D018105:DNA, Kinetoplast / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D007891:Leishmania / Q000235:genetics / Q000378:metabolism*
D000199:Actins; D002843:Chromatin; D043864:DNA, Catenated; D018105:DNA, Kinetoplast; D003850:Deoxyribonuclease I; D004250:DNA Topoisomerases, Type II