Capsid coding sequence is required for efficient replication of human rhinovirus 14 RNA

K L McKnight, S M Lemon
1996 Journal of Virology  
Mechanisms by which the plus-sense RNA genomes of picornaviruses are replicated remain poorly defined, but existing models do not suggest a role for sequences encoding the capsid proteins. However, candidate RNA replicons (⌬P1␤gal and ⌬P1Luc), representing the sequence of human rhinovirus 14 virus (HRV-14) with reporter protein sequences (␤-galactosidase or luciferase, respectively) replacing most of the P1 capsid-coding region, failed to replicate in transfected H1-HeLa cells despite efficient
more » ... s despite efficient primary cleavage of the polyprotein. To determine which P1 sequences might be required for RNA replication, HRV-14 mutants in which segments of the P1 region were removed in frame from the genome were constructed. Mutants with deletions involving the 5-proximal 1,489 nucleotides of the P1 region replicated efficiently, while those with deletions involving the 3 1,079 nucleotides did not. Reintroduction of the 3 P1 sequence into the nonreplicating ⌬P1Luc construct resulted in a new candidate replicon, ⌬P1Luc/VP3, which replicated well and expressed luciferase efficiently. Capsid proteins provided in trans by helper virus failed to rescue the nonreplicating ⌬P1Luc genome but were able to package the larger-than-genome-length ⌬P1Luc/VP3 replicon. Thus, a 3-distal P1 capsid-coding sequence has a previously unrecognized cis-active function related to replication of HRV-14 RNA.
doi:10.1128/jvi.70.3.1941-1952.1996 fatcat:gjuiw3ixp5efpelqate2atmr4u