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, J. Barklie Clements

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)33540-33549
Number of pages10
JournalJournal of Biological Chemistry
Volume278
Issue number35
DOIs
StatePublished - Aug 29 2003
Externally publishedYes

Fingerprint

Human Herpesvirus 1
Simplexvirus
Viruses
RNA
Proteins
Gene expression
Yeast
Yeasts
Poly G
RNA-Binding Proteins
Genomic Library
Herpesviridae
Viral DNA
Viral RNA
Viral Proteins
Life Cycle Stages
DNA Replication
Virion
Open Reading Frames
Nucleotides

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Identification of herpes simplex virus RNAs that interact specifically with regulatory protein ICP27 in vivo. / Sokolowski, Marcus; Scott, James E.; Heaney, Robert P.; Patel, Arvind H.; Clements, J. Barklie.

In: Journal of Biological Chemistry, Vol. 278, No. 35, 29.08.2003, p. 33540-33549.

Research output: Contribution to journalArticle

Sokolowski, Marcus ; Scott, James E. ; Heaney, Robert P. ; Patel, Arvind H. ; Clements, J. Barklie. / Identification of herpes simplex virus RNAs that interact specifically with regulatory protein ICP27 in vivo. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 35. pp. 33540-33549.
@article{e87365be2c07415d86dcd471bd191696,
title = "Identification of herpes simplex virus RNAs that interact specifically with regulatory protein ICP27 in vivo",
abstract = "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.",
author = "Marcus Sokolowski and Scott, {James E.} and Heaney, {Robert P.} and Patel, {Arvind H.} and Clements, {J. Barklie}",
year = "2003",
month = "8",
day = "29",
doi = "10.1074/jbc.M302063200",
language = "English",
volume = "278",
pages = "33540--33549",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "35",

}

TY - JOUR

T1 - Identification of herpes simplex virus RNAs that interact specifically with regulatory protein ICP27 in vivo

AU - Sokolowski, Marcus

AU - Scott, James E.

AU - Heaney, Robert P.

AU - Patel, Arvind H.

AU - Clements, J. Barklie

PY - 2003/8/29

Y1 - 2003/8/29

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0041355285&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0041355285&partnerID=8YFLogxK

U2 - 10.1074/jbc.M302063200

DO - 10.1074/jbc.M302063200

M3 - Article

VL - 278

SP - 33540

EP - 33549

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -