RNAs are packaged into human cytomegalovirus virions in proportion to their intracellular concentration. J Virol 2004 Oct;78(19):10390-8
Date
09/16/2004Pubmed ID
15367605Pubmed Central ID
PMC516422DOI
10.1128/JVI.78.19.10390-10398.2004Scopus ID
2-s2.0-4544262244 (requires institutional sign-in at Scopus site) 76 CitationsAbstract
The assembly of human cytomegalovirus (HCMV) virions is a complex process and involves the incorporation of viral transcripts. These RNAs are delivered to the newly infected cells and have the potential to be translated in the absence of HCMV gene expression. We have quantified the relative amount of RNAs in HCMV virions and infected cells with real-time reverse transcription-PCR and observed that viral and cellular RNAs are packaged in proportion to the amount of RNA within the cell at the time of assembly. To determine whether cis elements influenced RNA packaging, we constructed a recombinant HCMV mutant virus that expressed the yellow fluorescence protein (YFP) gene fused to the virion RNA UL21.5. We also constructed a mutant virus in which the UL21.5 transcription unit was replaced with the YFP gene. YFP RNA was incorporated into both viruses, indicating that RNA is incorporated in the absence of a virus-specific signal motif. Furthermore, with in situ hybridization, packaged transcripts were observed throughout the cytoplasm of the infected cells, including the site of virus assembly. Several proteins that nonspecifically interact with RNA, including the tegument protein pp28, were found within HCMV virions. These studies demonstrate that both viral and cellular RNAs are nonspecifically incorporated into HCMV, potentially through interactions with several virion proteins.
Author List
Terhune SS, Schröer J, Shenk TAuthor
Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Bacterial ProteinsCytomegalovirus
Genes, Reporter
Humans
Luminescent Proteins
RNA
RNA, Messenger
RNA, Viral
RNA-Binding Proteins
Viral Proteins
Virion
Virus Assembly