Originally published In Press as doi:10.1074/jbc.M302063200 on June 3, 2003
J. Biol. Chem., Vol. 278, Issue 35, 33540-33549, August 29, 2003
Identification of Herpes Simplex Virus RNAs That Interact Specifically with Regulatory Protein ICP27 in Vivo*
Marcus Sokolowski 
,
James E. Scott ,
Robert P. Heaney ,
Arvind H. Patel ¶ and
J. Barklie Clements ||
From the
Division of Virology, Institute of
Biomedical Life Sciences and ¶Medical Research
Council Virology Unit, Institute of Virology, Church Street, University of
Glasgow, Glasgow G11 5JR, Scotland, United Kingdom
Herpes simplex virus type 1 (HSV-1) protein ICP27 has an essential
regulatory role during viral replication, in part by post-transcriptional
control of gene expression, and has a counterpart in all herpes viruses
sequenced so far. Although much is known about the functions of this signature
herpesvirus protein, little is known about its RNA binding capabilities; ICP27
interacts with specificity for a subset of intronless HSV-1 RNAs and poly(G),
through its RGG box. We performed an in vivo yeast three-hybrid
screen of an HSV-1 genomic library, searching for ICP27 interacting RNAs.
Comparable with a yeast genomic screen, 24 of 55 single inserts mapped to
antisense strands of HSV-1 transcribed regions or non-transcribed regions. The
31 HSV-1 sense RNAs identified were 35 to 225 nucleotides in length and
interacted with preferred specificity for ICP27 as compared with an unrelated
RNA-binding protein. They map to 10 monocistronic and 10 polycistronic
transcripts of all kinetic classes and represent 28 open reading frames
encoding predominantly essential viral proteins with roles in viral DNA
replication and virion maturation. Several studies show regulatory effects by
ICP27 on the majority of these transcripts, consistent with its regulation of
the early-late switch in the HSV-1 life cycle. Deletion of the ICP27 RGG box
and the ICP27 M15 mutation, both lethal in virus, abolished or severely
reduced the ICP27-RNA interactions, indicating their biological relevance. The
study facilitates continued study of gene regulation by ICP27 by further
defining its interactions with viral RNAs.
Received for publication, February 27, 2003
, and in revised form, May 28, 2003.
* This work was supported by Medical Research Council funding (G9826324). The
costs of publication of this article were defrayed in part by the payment of
page charges. This article must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section 1734
solely to indicate this fact.
Present address: Capio Diagnostics, Mälarsjukhuset, SE-631 88
Eskilstuna, Sweden.
||
Recipient of a postdoctoral scholarship from the Swedish Foundation for
International Co-operation in Research and Higher Education. To whom
correspondence should be addressed: Inst. of Virology, University of Glasgow,
Church St., Glasgow G11 5JR, Scotland, UK. Tel.: 44-141-330-4037; Fax:
44-141-337-2236; E-mail:
b.clements{at}vir.gla.ac.uk.
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Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
