Study of yeast DNA topoisomerase II and its truncation derivatives by transmission electron microscopy.
The 1429-amino acid residue long yeast DNA topoisomerase II and three of its deletion derivatives, a C-terminal truncation containing residues 1-1202, a 92-kDa fragment spanning residues 410-1202, and an A'-fragment spanning residues 660-1202, were examined by transmission electron microscopy. Analysis of rotary-shadowed images of these molecules shows that the full-length enzyme assumes a tripartite structure, in which a large globular core comprising the carboxyl parts of the dimeric enzyme is connected to a pair of smaller spherical masses comprising the ATPase domains of the enzyme. The linkers bridging the large globular structure and each of the smaller spheres are not visible in most of the images but appear to be sufficiently stiff to keep the relative positions of the connected parts. The angle extended by the pair of spherical masses is variable and falls in a range of 50-100 degrees for the majority of the images. On binding of a nonhydrolyzable ATP analog to the enzyme, this angle is significantly reduced as the two spherical masses swing into contact. These observations, together with results from previous biochemical and x-ray crystallographic studies of the enzyme, provide a sketch of the molecular architecture and conformational states of a catalytically active type II DNA topoisomerase.
Benedetti
P
P
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, viale Marx 43, Rome 00137 Italy. piero@biocell.irmkant.rm.cnr.it
Silvestri
A
A
Fiorani
P
P
Wang
J C
JC
D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008961:Models, Structural; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D017384:Sequence Deletion
D010446:Peptide Fragments; D011994:Recombinant Proteins; D004250:DNA Topoisomerases, Type II