Structural Basis for the Recognition of Cellular mRNA Export Factor REF by Herpes Viral Proteins HSV-1 ICP27 and HVS ORF57
The herpesvirus proteins HSV-1 ICP27 and HVS ORF57 promote viral mRNA export by utilizing the cellular mRNA export machinery. This function is triggered by binding to proteins of the transcription-export (TREX) complex, in particular to REF/Aly which directs viral mRNA to the TAP/NFX1 pathway and, subsequently, to the nuclear pore for export to the cytoplasm. Here we have determined the structure of the REF-ICP27 interaction interface at atomic-resolution and provided a detailed comparison of the binding interfaces between ICP27, ORF57 and REF using solution-state NMR. Despite the absence of any obvious sequence similarity, both viral proteins bind on the same site of the folded RRM domain of REF, via short but specific recognition sites. The regions of ICP27 and ORF57 involved in binding by REF have been mapped as residues 104–112 and 103–120, respectively. We have identified the pattern of residues critical for REF/Aly recognition, common to both ICP27 and ORF57. The importance of the key amino acid residues within these binding sites was confirmed by site-directed mutagenesis. The functional significance of the ORF57-REF/Aly interaction was also probed using an ex vivo cytoplasmic viral mRNA accumulation assay and this revealed that mutants that reduce the protein-protein interaction dramatically decrease the ability of ORF57 to mediate the nuclear export of intronless viral mRNA. Together these data precisely map amino acid residues responsible for the direct interactions between viral adaptors and cellular REF/Aly and provide the first molecular details of how herpes viruses access the cellular mRNA export pathway.
Vyšlo v časopise:
Structural Basis for the Recognition of Cellular mRNA Export Factor REF by Herpes Viral Proteins HSV-1 ICP27 and HVS ORF57. PLoS Pathog 7(1): e32767. doi:10.1371/journal.ppat.1001244
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1001244
Souhrn
The herpesvirus proteins HSV-1 ICP27 and HVS ORF57 promote viral mRNA export by utilizing the cellular mRNA export machinery. This function is triggered by binding to proteins of the transcription-export (TREX) complex, in particular to REF/Aly which directs viral mRNA to the TAP/NFX1 pathway and, subsequently, to the nuclear pore for export to the cytoplasm. Here we have determined the structure of the REF-ICP27 interaction interface at atomic-resolution and provided a detailed comparison of the binding interfaces between ICP27, ORF57 and REF using solution-state NMR. Despite the absence of any obvious sequence similarity, both viral proteins bind on the same site of the folded RRM domain of REF, via short but specific recognition sites. The regions of ICP27 and ORF57 involved in binding by REF have been mapped as residues 104–112 and 103–120, respectively. We have identified the pattern of residues critical for REF/Aly recognition, common to both ICP27 and ORF57. The importance of the key amino acid residues within these binding sites was confirmed by site-directed mutagenesis. The functional significance of the ORF57-REF/Aly interaction was also probed using an ex vivo cytoplasmic viral mRNA accumulation assay and this revealed that mutants that reduce the protein-protein interaction dramatically decrease the ability of ORF57 to mediate the nuclear export of intronless viral mRNA. Together these data precisely map amino acid residues responsible for the direct interactions between viral adaptors and cellular REF/Aly and provide the first molecular details of how herpes viruses access the cellular mRNA export pathway.
Zdroje
1. HardyWR
Sandri-GoldinRM
1994 Herpes simplex virus inhibits host cell splicing, and regulatory protein ICP27 is required for this effect. J Virol 68 7790 7799
2. Sandri-GoldinRM
1998 ICP27 mediates HSV RNA export by shuttling through a leucine-rich nuclear export signal and binding viral intronless RNAs through an RGG motif. Genes Dev 12 868 879
3. Sandri-GoldinRM
2008 The many roles of the regulatory protein ICP27 during herpes simplex virus infection. Front Biosci 13 5241 5256
4. BoyneJR
ColganKJ
WhitehouseA
2008 Herpesvirus saimiri ORF57: a post-transcriptional regulatory protein. Front Biosci 13 2928 2938
5. BoyneJR
JacksonBR
TaylorA
MacnabSA
WhitehouseA
2010 Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs. EMBO J 29 1851 1864
6. StutzF
BachiA
DoerksT
BraunIC
SeraphinB
2000 REF, an evolutionary conserved family of hnRNP-like proteins, interacts with TAP/Mex67p and participates in mRNA nuclear export. RNA 6 638 650
7. CarmodySR
WenteSR
2009 mRNA nuclear export at a glance. J Cell Sci 122 1933 1937
8. KellySM
CorbettAH
2009 Messenger RNA export from the nucleus: a series of molecular wardrobe changes. Traffic 10 1199 1208
9. FribourgS
BraunIC
IzaurraldeE
ContiE
2001 Structural basis for the recognition of a nucleoporin FG repeat by the NTF2-like domain of the TAP/p15 mRNA nuclear export factor. Mol Cell 8 645 656
10. HautbergueGM
HungML
GolovanovAP
LianLY
WilsonSA
2008 Mutually exclusive interactions drive handover of mRNA from export adaptors to TAP. Proc Natl Acad Sci USA 105 5154 5159
11. KieslerE
MirallesF
VisaN
2002 HEL/UAP56 binds cotranscriptionally to the Balbiani ring pre-mRNA in an intron-independent manner and accompanies the BR mRNP to the nuclear pore. Curr Biol 12 859 862
12. SciabicaKS
DaiQJ
Sandri-GoldinRM
2003 ICP27 interacts with SRPK1 to mediate HSV splicing inhibition by altering SR protein phosphorylation. EMBO J 22 1608 1619
13. HautbergueGM
HungML
WalshMJ
SnijdersAP
ChangCT
2009 UIF, a New mRNA Export Adaptor that Works Together with REF/ALY, Requires FACT for Recruitment to mRNA. Curr Biol 19 1918 1924
14. JohnsonLA
LiL
Sandri-GoldinRM
2009 The cellular RNA export receptor TAP/NXF1 is required for ICP27-mediated export of herpes simplex virus 1 RNA, but the TREX complex adaptor protein Aly/REF appears to be dispensable. J Virol 83 6335 6346
15. StutzF
IzaurraldeE
2003 The interplay of nuclear mRNP assembly, mRNA surveillance and export. Trends Cell Biol 13 319 327
16. PhelanA
ClementsJB
1997 Herpes simplex virus type 1 immediate early protein IE63 shuttles between nuclear compartments and the cytoplasm. J Gen Virol 78 Pt 12 3327 3331
17. ChenIH
LiL
SilvaL
Sandri-GoldinRM
2005 ICP27 recruits Aly/REF but not TAP/NXF1 to herpes simplex virus type 1 transcription sites although TAP/NXF1 is required for ICP27 export. J Virol 79 3949 3961
18. JohnsonLA
Sandri-GoldinRM
2009 Efficient nuclear export of herpes simplex virus 1 transcripts requires both RNA binding by ICP27 and ICP27 interaction with TAP/NXF1. J Virol 83 1184 1192
19. KoffaMD
ClementsJB
IzaurraldeE
WaddS
WilsonSA
2001 Herpes simplex virus ICP27 protein provides viral mRNAs with access to the cellular mRNA export pathway. EMBO J 20 5769 5778
20. WhitehouseA
CooperM
MeredithDM
1998 The Immediate-Early gene product encoded by open reading frame 57 of Herpesvirus Saimiri modulates gene expression at a posttranscriptional level. J Virol 72 857 861
21. GoodwinDJ
WhitehouseA
2001 A gamma-2 herpesvirus nucleocytoplasmic shuttle protein interacts with importin alpha 1 and alpha 5. J Biol Chem 276 19905 19912
22. WilliamsBJ
BoyneJR
GoodwinDJ
RoadenL
HautbergueGM
2005 The prototype gamma-2 herpesvirus nucleocytoplasmic shuttling protein, ORF 57, transports viral RNA through the cellular mRNA export pathway. Biochem J 387 295 308
23. ChenIH
SciabicaKS
Sandri-GoldinRM
2002 ICP27 interacts with the RNA export factor Aly/REF to direct herpes simplex virus type 1 intronless mRNAs to the TAP export pathway. J Virol 76 12877 12889
24. MearsWE
RiceSA
1996 The RGG box motif of the herpes simplex virus ICP27 protein mediates an RNA-binding activity and determines in vivo methylation. J Virol 70 7445 7453
25. LengyelJ
GuyC
LeongV
BorgeS
RiceSA
2002 Mapping of functional regions in the amino-terminal portion of the herpes simplex virus ICP27 regulatory protein: importance of the leucine-rich nuclear export signal and RGG Box RNA-binding domain. J Virol 76 11866 11879
26. YuJ
ShinB
ParkES
YangS
ChoiS
2010 Protein arginine methyltransferase 1 regulates herpes simplex virus replication through ICP27 RGG-box methylation. Biochem Biophys Res Commun 391 322 328
27. SoukiSK
Sandri-GoldinRM
2009 Arginine methylation of the ICP27 RGG box regulates the functional interactions of ICP27 with SRPK1 and Aly/REF during herpes simplex virus 1 infection. J Virol 83 8970 8975
28. GoodwinDJ
HallKT
StevensonAJ
MarkhamAF
WhitehouseA
1999 The open reading frame 57 gene product of herpesvirus saimiri shuttles between the nucleus and cytoplasm and is involved in viral RNA nuclear export. J Virol 73 10519 10524
29. ColganKJ
BoyneJR
WhitehouseA
2009 Identification of a response element in a herpesvirus saimiri mRNA recognized by the ORF57 protein. J Gen Virol 90 596 601
30. HiriartE
FarjotG
GruffatH
NguyenMV
SergeantA
2003 A novel nuclear export signal and a REF interaction domain both promote mRNA export by the Epstein-Barr virus EB2 protein. J Biol Chem 278 335 342
31. MalikP
BlackbournDJ
ClementsJB
2004 The evolutionarily conserved Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with REF protein and acts as an RNA export factor. J Biol Chem 279 33001 33011
32. Perez-AlvaradoGC
Martinez-YamoutM
AllenMM
GrosschedlR
DysonHJ
2003 Structure of the nuclear factor ALY: insights into post-transcriptional regulatory and mRNA nuclear export processes. Biochemistry 42 7348 7357
33. GolovanovAP
HautbergueGM
TintaruAM
LianLY
WilsonSA
2006 The solution structure of REF2-I reveals interdomain interactions and regions involved in binding mRNA export factors and RNA. RNA 12 1933 1948
34. GolovanovAP
HautbergueGM
WilsonSA
LianLY
2006 Assignment of 1H, 13C, and 15N resonances for the REF2-I mRNA export factor. J Biomol NMR 36 Suppl 1 41
35. BoyneJR
ColganKJ
WhitehouseA
2008 Recruitment of the complete hTREX complex is required for Kaposi's sarcoma-associated herpesvirus intronless mRNA nuclear export and virus replication. PLoS Pathog 4 e1000194
36. BoyneJR
WhitehouseA
2009 Nucleolar disruption impairs Kaposi's sarcoma-associated herpesvirus ORF57-mediated nuclear export of intronless viral mRNAs. FEBS Lett 583 3549 3556
37. TintaruAM
HautbergueGM
HounslowAM
HungML
LianLY
2007 Structural and functional analysis of RNA and TAP binding to SF2/ASF. EMBO Rep 8 756 762
38. HargousY
HautbergueGM
TintaruAM
SkrisovskaL
GolovanovAP
2006 Molecular basis of RNA recognition and TAP binding by the SR proteins SRp20 and 9G8. EMBO J 25 5126 5137
39. Corbin-LickfettKA
RojasS
LiL
CoccoMJ
Sandri-GoldinRM
2010 ICP27 Phosphorylation Site Mutants Display Altered Functional Interactions with Cellular Export Factors Aly/REF and TAP/NXF1 but Are Able to Bind Herpes Simplex Virus 1 RNA. J Virol 84 2212 2222
40. ColganKJ
BoyneJR
WhitehouseA
2009 Uncoupling of hTREX demonstrates that UAP56 and hTHO-complex recruitment onto herpesvirus saimiri intronless transcripts is required for replication. J Gen Virol 90 1455 1460
41. MearsWE
LamV
RiceSA
1995 Identification of nuclear and nucleolar localization signals in the herpes simplex virus regulatory protein ICP27. J Virol 69 935 947
42. KielkopfCL
RodionovaNA
GreenMR
BurleySK
2001 A novel peptide recognition mode revealed by the X-ray structure of a core U2AF-(35)/U2AF(65) heterodimer. Cell 106 595 605
43. KielkopfCL
LuckeS
GreenAR
2004 U2AF homology motifs: protein recognition in the RRM world. Genes & Development 18 1513 1526
44. CorsiniL
BonnalS
BasquinJ
HothornM
ScheffzekK
2007 U2AF-homology motif interactions are required for alternative splicing regulation by SPF45. Nature Structural & Molecular Biology 14 620 629
45. RideauAP
GoodingC
SimpsonPJ
MonieTP
LorenzM
2006 A peptide motif in Raver1 mediates splicing repression by interaction with the PTB RRM2 domain. Nature Structural & Molecular Biology 13 839 848
46. CleryA
BlatterM
AllainFH
2008 RNA recognition motifs: boring? Not quite. Curr Opin Struct Biol 18 290 298
47. GouldF
HarrisonSM
HewittEW
WhitehouseA
2009 Kaposi's sarcoma-associated herpesvirus RTA promotes degradation of the Hey1 repressor protein through the ubiquitin proteasome pathway. J Virol 83 6727 6738
48. HallKT
GilesMS
CalderwoodMA
GoodwinDJ
MatthewsDA
2002 The Herpesvirus Saimiri open reading frame 73 gene product interacts with the cellular protein p32. J Virol 76 11612 11622
49. BoyneJR
WhitehouseA
2006 Nucleolar trafficking is essential for nuclear export of intronless herpesvirus mRNA. Proc Natl Acad Sci USA 103 15190 15195
50. DelaglioF
GrzesiekS
VuisterGW
ZhuG
PfeiferJ
1995 NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6 277 293
51. CornilescuG
DelaglioF
BaxA
1999 Protein backbone angle restraints from searching a database for chemical shift and sequence homology. J Biomol NMR 13 289 302
52. GuntertP
2004 Automated NMR structure calculation with CYANA. Methods Mol Biol 278 353 378
53. ZwahlenC
LegaultP
VincentSJF
GreenblattJ
KonratR
1997 Methods for Measurement of Intermolecular NOEs by Multinuclear NMR Spectroscopy: Application to a Bacteriophage l N-Peptide/boxB RNA Complex. J Am Chem Soc 119 6711 6721
54. StuartAC
BorzilleriKA
WithkaJM
PalmerAG
1999 Compensating for Variations in 1H-13C Scalar Coupling Constants in Isotope-Filtered NMR Experiments. J Am Chem Soc 121 5346 5347
55. CavanaghJ
FairbrotherWJ
PalmerAGr
RanceM
SkeltonNJ
2007 Protein NMR Spectroscopy: Principles and Practice. London, UK Academic Press
56. GuexN
PeitschMC
1997 SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18 2714 2723
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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