Morphogenesis of human adenovirus type 2: Sequence of entry of proteins into previral and viral particles

Abstract

The initial steps of adenovirus capsid morphogenesis and the sequence of entry of structural and nonstructural proteins into assembly-intermediate (IM) particles were investigated by pulse-chase labeling, temperature shifts, and cycloheximide inhibition of particle formation. The experiments were performed on wild-type and two assembly-defective, temperature-sensitive mutants, H2 ts 112 and H2 ts 107. The sequence of events in the adenovirus assembly can be schematized as follows. (i) Hexons, pentons, and protein IX assembled with scaffolding proteins 100K, PVIII, and PVII, precursor to the major core protein, to form a previral particle banding at a density of 1.285 in CsCl; (ii) additional incorporation of maturation and/or stabilization proteins IIIa, 50K, 39K, 28K, and PVI led to 1.295 IM; (iii) exit of 100K, 39K, and 28K, and entry of viral DNA gave rise to 1.370 IM; (iv) dephosphorylation and/or exit of 50K and exchange with core protein V and processing of precursors to VII, VI, VIII, and DNA-terminal protein resulted in formation of infectious 1.345 virion. The polypeptide composition of the new class of assembly-intermediate particles elicited by H2 is 107 (1.285 IM), suggested that 100K, PVIII, and also PVII might serve as scaffold components for adenovirus capsid building.