Retroviral capsid assembly: a role for the CA dimer in initiation.
In maturing retroviral virions, CA protein assembles to form a capsid shell that is essential for infectivity. The structure of the two folded domains [N-terminal domain (NTD) and C-terminal domain (CTD)] of CA is highly conserved among various retroviruses, and the capsid assembly pathway, although poorly understood, is thought to be conserved as well. In vitro assembly reactions with purified CA proteins of the Rous sarcoma virus (RSV) were used to define factors that influence the kinetics of capsid assembly and provide insights into underlying mechanisms. CA multimerization was triggered by multivalent anions providing evidence that in vitro assembly is an electrostatically controlled process. In the case of RSV, in vitro assembly was a well-behaved nucleation-driven process that led to the formation of structures with morphologies similar to those found in virions. Isolated RSV dimers, when mixed with monomeric protein, acted as efficient seeds for assembly, eliminating the lag phase characteristic of a monomer-only reaction. This demonstrates for the first time the purification of an intermediate on the assembly pathway. Differences in the intrinsic tryptophan fluorescence of monomeric protein and the assembly-competent dimer fraction suggest the involvement of the NTD in the formation of the functional dimer. Furthermore, in vitro analysis of well-characterized CTD mutants provides evidence for assembly dependence on the second domain and suggests that the establishment of an NTD-CTD interface is a critical step in capsid assembly initiation. Overall, the data provide clear support for a model whereby capsid assembly within the maturing virion is dependent on the formation of a specific nucleating complex that involves a CA dimer and is directed by additional virion constituents.
Purdy
John G
JG
Department of Microbiology and Immunology, College of Medicine, The Pennsylvania State University, 500 University Drive, Hershey, PA 17033, USA.
Flanagan
John M
JM
Ropson
Ira J
IJ
Craven
Rebecca C
RC
D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins / Q000502:physiology*; D007700:Kinetics; D055503:Protein Multimerization; D012190:Retroviridae; D052736:Rous sarcoma virus / Q000502:physiology*; D055672:Static Electricity; D019065:Virus Assembly
D036022:Capsid Proteins