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J Virol, May 1998, p. 3837-3844, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Temporal Mapping of Transcripts in Herpesvirus 6 Variants

Prisco Mirandola, Paola Menegazzi, Stefania Merighi, Tullia Ravaioli, Enzo Cassai, and Dario Di Luca*

Dipartimento di Medicina Sperimentale e Diagnostica, Sezione di Microbiologia, Università di Ferrara, Ferrara, Italy

Received 22 December 1997/Accepted 30 January 1998

To define the molecular features characteristic of the early stages of infection of lymphocytes with human herpesvirus 6 (HHV-6) variant A or B, we studied the temporal regulation of expression of selected sets of viral genes. Thus, U42, U94, U89-U90, U73, and U39 are alpha  genes since their transcripts (i) were made in the presence of inhibitors of protein synthesis and (ii) were detected 3 h after infection of untreated cells. U41, U53, U31, and U19 are beta  genes since their expression is inhibited by cycloheximide but not by phosphonoacetate, an inhibitor of DNA synthesis. U100 is a gamma  gene since its spliced transcript encoding the structural glycoprotein gp82/105 was first detected 16 h after infection of untreated cells but could not be detected in cells treated with phosphonoacetate. HHV-6 variants differ in the transcription patterns of their genes. U16-U17 originates a splice transcript and is regulated as alpha  in HHV-6B and as beta  in HHV-6A. U91 generates two transcripts, amplified as 476- and 374-bp PCR fragments. The 476-bp fragment is alpha  in HHV-6A-infected cells but beta  in HHV-6B-infected cells. Conversely, the 374-bp fragment is beta  in HHV-6A-infected cells and alpha  in HHV-6B-infected cells. Furthermore, the spliced product of U18-U19-U20 (526 bp) is beta  in HHV-6A-infected cells, but only a partially spliced form (1.9 kb) was detected at late stages of infection in HHV-6B. HHV-6 transcription was also studied in nonproductive lymphoid cells, and the same transcription pattern detected during lytic infection was observed. Also, HHV-6 variants maintain the differences in U91, U16-17, and U18-U19-U20. We conclude that, as expected from the sequencing data, gene expression is generally similar in HHV-6 variants. However, transcription of selected genes in HHV-6A and HHV-6B differs with respect to temporal regulation and splicing pattern. Furthermore, the identification of viral functions expressed during the different stages of lytic replication suggests that reverse transcription-PCR for HHV-6 genes is a useful diagnostic approach to differentiate between latent and productive HHV-6 infection.

* Corresponding author. Mailing address: Sezione di Microbiologia, Dipartimento di Medicina Sperimentale e Diagnostica, Università di Ferrara, Via Borsari 46, 44100 Ferrara, Italy. Phone: (39) 532 291408. Fax: (39) 532 247618. E-mail: dil{at}dns.unife.it.

J Virol, May 1998, p. 3837-3844, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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