Abstract
The RNA-dependent protein kinase (PKR) is an interferon-induced, RNA-activated enzyme that phosphorylates the eukaryotic initiation factor 2α, rendering the translation machinery inactive. Viruses have developed strategies for preventing the action of PKR, one of which is the production of small RNAs that inhibit the enzyme. Epstein-Barr virus (EBV) encodes EBER1, a 167 nucleotide non-coding RNA that is constitutively expressed by the EBV-infected cells. EBER1 binds PKR in vitro and has been shown to prevent inhibition of translation by PKR in vitro. We used affinity cleavage by the EDTA·Fe-modified double-stranded RNA-binding domain (dsRBD) of PKR to show that stem-loop IV (nucleotides 87-123) of EBER1 makes specific contacts with the dsRBD. To further demonstrate the specificity of this interaction, we generated a deletion mutant of EBER1, comprising only stem-loop IV (mEBER1). Cleavage patterns produced on mEBER1 by the bound dsRBD were remarkably similar to those found on full-length EBER1. Using cleavage data from two different dsRBD mutants, we present a model of the interaction of PKR dsRBD and mEBER1.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 622-627 |
| Number of pages | 6 |
| Journal | EMBO Reports |
| Volume | 3 |
| Issue number | 7 |
| DOIs | |
| State | Published - 2002 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
- Genetics
Cite this
The binding site of the RNA-dependent protein kinase (PKR) on EBER1 RNA from Epstein-Barr virus. / Vuyisich, Momchilo; Spanggord, Richard J.; Beal, Peter A.
In: EMBO Reports, Vol. 3, No. 7, 2002, p. 622-627.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The binding site of the RNA-dependent protein kinase (PKR) on EBER1 RNA from Epstein-Barr virus
AU - Vuyisich, Momchilo
AU - Spanggord, Richard J.
AU - Beal, Peter A.
PY - 2002
Y1 - 2002
N2 - The RNA-dependent protein kinase (PKR) is an interferon-induced, RNA-activated enzyme that phosphorylates the eukaryotic initiation factor 2α, rendering the translation machinery inactive. Viruses have developed strategies for preventing the action of PKR, one of which is the production of small RNAs that inhibit the enzyme. Epstein-Barr virus (EBV) encodes EBER1, a 167 nucleotide non-coding RNA that is constitutively expressed by the EBV-infected cells. EBER1 binds PKR in vitro and has been shown to prevent inhibition of translation by PKR in vitro. We used affinity cleavage by the EDTA·Fe-modified double-stranded RNA-binding domain (dsRBD) of PKR to show that stem-loop IV (nucleotides 87-123) of EBER1 makes specific contacts with the dsRBD. To further demonstrate the specificity of this interaction, we generated a deletion mutant of EBER1, comprising only stem-loop IV (mEBER1). Cleavage patterns produced on mEBER1 by the bound dsRBD were remarkably similar to those found on full-length EBER1. Using cleavage data from two different dsRBD mutants, we present a model of the interaction of PKR dsRBD and mEBER1.
AB - The RNA-dependent protein kinase (PKR) is an interferon-induced, RNA-activated enzyme that phosphorylates the eukaryotic initiation factor 2α, rendering the translation machinery inactive. Viruses have developed strategies for preventing the action of PKR, one of which is the production of small RNAs that inhibit the enzyme. Epstein-Barr virus (EBV) encodes EBER1, a 167 nucleotide non-coding RNA that is constitutively expressed by the EBV-infected cells. EBER1 binds PKR in vitro and has been shown to prevent inhibition of translation by PKR in vitro. We used affinity cleavage by the EDTA·Fe-modified double-stranded RNA-binding domain (dsRBD) of PKR to show that stem-loop IV (nucleotides 87-123) of EBER1 makes specific contacts with the dsRBD. To further demonstrate the specificity of this interaction, we generated a deletion mutant of EBER1, comprising only stem-loop IV (mEBER1). Cleavage patterns produced on mEBER1 by the bound dsRBD were remarkably similar to those found on full-length EBER1. Using cleavage data from two different dsRBD mutants, we present a model of the interaction of PKR dsRBD and mEBER1.
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UR - http://www.scopus.com/inward/citedby.url?scp=0036023940&partnerID=8YFLogxK
U2 - 10.1093/embo-reports/kvf137
DO - 10.1093/embo-reports/kvf137
M3 - Article
VL - 3
SP - 622
EP - 627
JO - EMBO Reports
JF - EMBO Reports
SN - 1469-221X
IS - 7
ER -