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Journal of Virology, May 2001, p. 4376-4385, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4376-4385.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Herpes Simplex Virus IE63 (ICP27) Protein Interacts with Spliceosome-Associated Protein 145 and Inhibits Splicing prior to the First Catalytic Step

H. E. Bryant,1,dagger S. E. Wadd,1 A. I. Lamond,2 S. J. Silverstein,3 and J. B. Clements1,*

Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR,1 and Division of Gene Regulation and Expression, The Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH,2 Scotland, United Kingdom, and Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 100323

Received 6 November 2000/Accepted 6 February 2001

The multifunctional herpes simplex virus type 1 (HSV-1) protein IE63 (ICP27) interacts with the essential pre-mRNA splicing factor, spliceosome-associated protein 145 (SAP145), and in infected cells IE63 and SAP145 colocalize. This interaction was reduced or abrogated completely using extracts from cells infected with IE63 viral mutants, with mutations in IE63 KH and Sm homology domains, which do not exhibit host shutoff or inhibit splicing. In the presence of IE63, splicing in vitro was inhibited prior to the first catalytic step and the B/C complex formed during splicing was shifted up in mobility and reduced in intensity. With the use of splicing extracts, IE63 and SAP145 both comigrated with the B/C complex, suggesting that they interact within this complex to inhibit B/C complex formation or conversion. The inhibition of splicing may facilitate the export of viral or cellular transcripts, possibly via other protein partners of IE63. These data provide important new insights into how IE63 influences pre-mRNA processing during HSV-1 infection.

* Corresponding author. Mailing address: Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Church St., Glasgow G11 5JR, Scotland, United Kingdom. Phone: 44 141 330 4027. Fax: 44 141 337 2236. E-mail: b.clements{at}vir.gla.ac.uk.

dagger Present address: Ludwig Institute for Cancer Research, St. Mary's Hospital Medical School, London W2 1PG, United Kingdom.

Journal of Virology, May 2001, p. 4376-4385, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4376-4385.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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