On the association of DNA polymerase alpha activity with the nuclear matrix in HeLa cells.
The association of DNA polymerase alpha activity with the nuclear matrix has been reinvestigated in HeLa cells. Isolated nuclei were extracted with 2M NaCl and then digested with Dnase I and the final structures were recovered by centrifugation through a sucrose cushion. Typically over 98% of the total DNA synthesized in the matrix fraction on either endogenous matrix-associated DNA or activated calf thymus DNA was due to DNA polymerase alpha as defined by inhibition to n-ethylmaleimide or aphidicolin. DNA polymerase beta activity was absent or recovered in only trace amounts. Matrix-bound DNA polymerase alpha activity demonstrated a remarkable degree of stability: DNA synthesis was essentially linear up to 3 hours at 37 degrees C. Overall, these results substantiate previous findings from regenerating rat liver, unlike other data obtained from tissue culture cells.
Martelli
A M
AM
Istituto di Anatomia Umana Normale, Bologna, Italy.
Cocco
L
L
Manzoli
F A
FA
D016590:Aphidicolin; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D004256:DNA Polymerase I / Q000378:metabolism; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004261:DNA Replication / Q000502:physiology*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004224:Diterpenes / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D015530:Nuclear Matrix / Q000201:enzymology* / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism; D013696:Temperature
D004224:Diterpenes; D016590:Aphidicolin; D012965:Sodium Chloride; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D003850:Deoxyribonuclease I; D005033:Ethylmaleimide