Topoisomerase II forms multimers in vitro: effects of metals, beta-glycerophosphate, and phosphorylation of its C-terminal domain.
We present a novel assay for the study of protein-protein interactions involving DNA topoisomerase II. Under various conditions of incubation we observe that topoisomerase II forms complexes at least tetrameric in size, which can be sedimented by centrifugation through glycerol. The multimers are enzymatically active and can be visualized by electron microscopy. Dephosphorylation of topoisomerase II inhibits its multimerization, which can be restored at least partially by rephosphorylation of multiple sites within its 200 C-terminal amino acids by casein kinase II. Truncation of topoisomerase II just upstream of the major phosphoacceptor sites reduces its aggregation, rendering the truncated enzyme insensitive to either kinase treatments or phosphatase treatments. This is consistent with a model in which interactions involving the phosphorylated C-terminal domain of topoisomerase II aid either in chromosome segregation or in chromosome condensation.
Vassetzky
Y S
YS
Swiss Institute for Experimental Cancer Research, Epalinges.
Dang
Q
Q
Benedetti
P
P
Gasser
S M
SM
D002118:Calcium / Q000494:pharmacology; D002875:Chromosomes / Q000648:ultrastructure; D003300:Copper / Q000494:pharmacology; D004247:DNA / Q000378:metabolism / Q000648:ultrastructure; D004250:DNA Topoisomerases, Type II / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005994:Glycerophosphates / Q000494:pharmacology; D007399:Interphase / Q000502:physiology; D008854:Microscopy, Electron; D009687:Nuclear Proteins / Q000378:metabolism; D009711:Nucleotides / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D004268:DNA-Binding Proteins; D005994:Glycerophosphates; D009687:Nuclear Proteins; D009711:Nucleotides; D010446:Peptide Fragments; D003300:Copper; D004247:DNA; D004250:DNA Topoisomerases, Type II; D002118:Calcium; C031463:beta-glycerophosphoric acid