Treatment with a polyamine analog alters DNA-matrix association in HeLa cell nuclei: a nucleoid halo assay.
The polyamine analog 1,14-bis(ethylamino)-5,10-diazatetradecane (BE-4-4-4) depletes polyamines and inhibits the growth of tumor cells in tissue culture. We treated HeLa cells in culture with BE-4-4-4 for different time periods to produce different degrees of polyamine depletion. The cells were lysed and dehistonized to obtain nucleoids containing DNA attached to the nuclear matrix. Titration of the nucleoids with propidium iodide caused an uncoiling of negatively supercoiled DNA, resulting in the formation of a halo surrounding the nucleoid periphery. The halo diameters in both the BE-4-4-4-treated cells and the untreated control cells were measured using a fluorescence image analysis system. As compared to the control cells, the BE-4-4-4-treated cells showed a 20-25% decrease in halo diameter, indicating that there was less relaxation of the negative supercoils in the nuclear DNA of the BE-4-4-4-treated cells than in the controls.
Basu
H S
HS
Department of Neurological Surgery, University of California, San Francisco 94143.
Wright
W D
WD
Deen
D F
DF
Roti-Roti
J
J
Marton
L J
LJ
D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D015530:Nuclear Matrix / Q000187:drug effects / Q000378:metabolism; D011073:Polyamines / Q000378:metabolism*; D013096:Spermine / Q000031:analogs & derivatives* / Q000378:metabolism
D004273:DNA, Neoplasm; D006657:Histones; D011073:Polyamines; C058306:N(1),N(14)-bis(ethyl)homospermine; D013096:Spermine