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Journal of Virology, August 2000, p. 7307-7319, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Processing of
-Globin and ICP0 mRNA in Cells
Infected with Herpes Simplex Virus Type 1 ICP27 Mutants
Kimberly S.
Ellison,1
Stephen A.
Rice,2
Robert
Verity,1 and
James R.
Smiley1,*
Department of Medical Microbiology and
Immunology, University of Alberta, Edmonton, Alberta, Canada T6G
2H7,1 and Department of Microbiology,
University of Minnesota Medical School, Minneapolis, Minnesota
554552
Received 2 March 2000/Accepted 17 May 2000
Herpes simplex virus (HSV) ICP27 is an essential and
multifunctional regulator of viral gene expression that modulates RNA splicing, polyadenylation, and nuclear export. We have previously reported that ICP27 causes the cytoplasmic accumulation of unspliced
-globin pre-mRNA. Here we examined the effects of a series of ICP27
mutations that alter important functional regions of the protein on the
processing and nuclear transport of
-globin and HSV ICP0 RNA. The
results demonstrate that ICP27 mutants that are impaired for growth in
noncomplementing cells, including mutants in the N- and C-terminal
regions, are defective in the accumulation of
-globin pre-mRNA.
Unexpectedly, several mutants that are competent to repress the
expression of reporter genes in transient transfection assays failed to
accumulate unspliced RNA, implying that different mechanisms are
responsible for transrepression and pre-mRNA accumulation. Several
mutants caused a marked increase in the length and heterogeneity of the
-globin mRNA poly(A) tail, suggesting that ICP27 may directly or
indirectly affect the regulation of poly(A) polymerase. ICP27 was also
required for the accumulation of multiple ICP0 intron-bearing transcripts, but this effect displayed a mutational sensitivity profile
different from that of accumulation of unspliced
-globin RNA.
Moreover, unlike spliced and unspliced
-globin RNAs, which were
efficiently exported to the cytoplasm, spliced and intron-containing ICP0 transcripts were predominantly nuclear in localization, and ICP27
was not required for nuclear retention of the spliced message. We
propose that these transcript- and ICP27 allele-specific differences may be explained by the presence of a strong cis-acting
ICP27 response element in the
-globin transcript.
*
Corresponding author. Mailing address: Department of
Medical Microbiology & Immunology, 1-41 Medical Sciences Bldg.,
University of Alberta, Edmonton, Alberta, Canada T6G 2H7. Phone: (403)
492-2308. Fax: (403) 492-7521. E-mail:
jim.smiley{at}ualberta.ca.
Journal of Virology, August 2000, p. 7307-7319, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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