Structural basis for the cooperative assembly of large T antigen on the origin of replication.
Large T antigen (LTag) from simian virus 40 (SV40) is an ATP-driven DNA helicase that specifically recognizes the core of the viral origin of replication (ori), where it oligomerizes as a double hexamer. During this process, binding of the first hexamer stimulates the assembly of a second one. Using electron microscopy, we show that the N-terminal part of LTag that includes the origin-binding domain does not present a stable quaternary structure in single hexamers. This disordered region, however, is well arranged within the LTag double hexamer after specific ori recognition, where it mediates the interactions between hexamers and constructs a separated structural module at their junction. We conclude that full assembly of LTag hexamers occurs only within the dodecamer, and requires the specific hexamer-hexamer interactions established upon binding to the origin of replication. This mechanism provides the structural basis for the cooperative assembly of LTag double hexamer on the cognate viral ori.
Centro Nacional de Biotecnología, Darwin 3, Cantoblanco 28049 Madrid, Spain.
Mikel
M
Valle
Xiaojiang S
XS
Chen
Luis Enrique
LE
Donate
Ellen
E
Fanning
José María
JM
Carazo
D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry* / Q000378:metabolism; D020285:Cryoelectron Microscopy; D004279:DNA, Viral; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D018741:Replication Origin; D013539:Simian virus 40
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral