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Journal of Virology, December 1998, p. 9889-9896, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Splicing-Independent Expression of the Herpes Simplex Virus Type 1 Thymidine Kinase Gene Is Mediated by Three cis-Acting RNA Subelements

Glen C. Otero and Thomas J. Hope*

Infectious Disease Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037

Received 11 May 1998/Accepted 27 August 1998

Herpes simplex virus genes are predominantly intronless. We identified cis-acting elements in the intronless herpes simplex virus type 1 thymidine kinase (TK) gene that facilitate intron-independent gene expression. TK sequences functionally replaced the hepatitis B virus (HBV) posttranscriptional regulatory element (PRE) by inducing the expression of the intronless HBV surface message. TK also activated the pDM138 assay by inducing the cytoplasmic accumulation of intron-containing RNA. Multiple cis-acting RNA sequences, or subelements, that induce cytoplasmic localization of unspliced RNA were mapped within the TK gene. The presence of multiple RNA subelements within the TK gene is reminiscent of the multiple subelements in the HBV PRE required for the cytoplasmic accumulation of intronless HBV RNAs. Similar to HBV PRE subelements, duplication of a single TK subelement resulted in greater-than-additive increases in activity. A reporter chimera containing a single TK subelement juxtaposed to an HBV PRE subelement demonstrated a commensurate increase in activity. These results suggest that viral intronless genes utilize a similar strategy for intron-independent gene expression that requires multiple cis-acting RNA signals. Furthermore, like HBV PRE-containing RNA, TK cytoplasmic localization is not sensitive to leptomycin B, a drug that inhibits the export of proteins containing nuclear export signals. From this, we conclude that proteins that bind TK and facilitate its cytoplasmic accumulation do not travel through a CRM1-dependent RNA transport pathway.

* Corresponding author. Mailing address: The Salk Institute for Biological Studies, Infectious Disease Laboratory, P.O. Box 85800, La Jolla, CA 92037. Phone: (619) 453-4100, ext. 1559. Fax: (619) 554-0341. E-mail: Hope{at}salk.edu.

Journal of Virology, December 1998, p. 9889-9896, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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