The extreme carboxyl terminus of the equine herpesvirus 1 homolog of herpes simplex virus VP16 is essential for immediate-early gene activation.
Abstract: Gene 12 of equine herpesvirus 1 (EHV-1), the homolog of herpes simplex virus (HSV) VP16 (alpha TIF, Vmw65), was cloned into a eukaryotic expression vector by PCR and used in transactivation studies of both the EHV-1 and HSV-1 IE1 promoters. Results demonstrated that the product of gene 12 is a potent transactivator of immediate-early gene expression of both viruses, which requires sequences in the upstream HSV-1 promoter for activity. Mutational analysis of the gene 12 open reading frame indicated that removal of the C-terminal 7 amino acids, which contain a short region of homology with the extreme C terminus of VP16, inactivated the protein. Within this region, only a single methionine residue appeared to be essential for activity, implying that gene 12 may have a modular array of organization similar to that of VP16. However, fusion of the gene 12 C terminus to a truncated form of VP16, which contained the complex formation domain, did not restore activity to the HSV-1 protein. These data demonstrate that the EHV-1 immediate-early transactivator may not be functionally colinear with VP16, with transactivation requiring both the C terminus and another region(s) present within the N-terminal portion.
Publication Date: 1994-08-01 PubMed ID: 8035487PubMed Central: PMC236429DOI: 10.1128/JVI.68.8.4890-4897.1994Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research paper discusses a study on the gene 12 of equine herpesvirus 1 (EHV-1) and its role in the immediate-early gene expression of both EHV-1 and herpes simplex virus (HSV-1), as well as the importance of a single methionine residue in the C-terminal region of this gene.
Study on Gene 12 of Equine Herpesvirus 1 (EHV-1)
- The researchers cloned gene 12 of EHV-1, a homolog of herpes simplex virus (HSV) VP16, into a eukaryotic expression vector using PCR.
- The goal was to use this cloning in transactivation studies of the EHV-1 and HSV-1 IE1 promoters.
- The finding revealed that the product of gene 12 acts as a potent transactivator of immediate-early gene expression for both viruses.
- It was found that the activity of this gene product requires sequences in the upstream HSV-1 promoter.
Mutational Analysis of Gene 12
- The researchers carried out a mutational analysis on the open reading frame of gene 12.
- They found that by removing the C-terminal 7 amino acids, which contain a short region of homology with the extreme C terminus of VP16, resulted in the inactivation of the protein.
- Within this region, only a single methionine residue appeared to be essential for activity, suggesting that gene 12 may be organized similarly to VP16.
Comparing Functionality of EHV-1 with VP16
- In the study, it was observed that mere fusion of the gene 12 C terminus to a truncated form of VP16 did not restore activity to the HSV-1 protein.
- This implies that the EHV-1 immediate-early transactivator might not be functionally equivalent to VP16.
- Transactivation requires both the C terminus and another region(s) present within the N-terminal portion of the molecular structure.
Cite This Article
APA
Elliott GD.
(1994).
The extreme carboxyl terminus of the equine herpesvirus 1 homolog of herpes simplex virus VP16 is essential for immediate-early gene activation.
J Virol, 68(8), 4890-4897.
https://doi.org/10.1128/JVI.68.8.4890-4897.1994 Publication
Researcher Affiliations
- Department of Microbiology, University of Leeds, United Kingdom.
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Cell Line
- Cloning, Molecular
- DNA, Viral
- Gene Expression Regulation, Viral
- Herpes Simplex Virus Protein Vmw65 / chemistry
- Herpes Simplex Virus Protein Vmw65 / genetics
- Herpesvirus 1, Equid / genetics
- Immediate-Early Proteins / genetics
- Methionine / metabolism
- Molecular Sequence Data
- Promoter Regions, Genetic
- Sequence Homology
- Transcriptional Activation
- Vero Cells
Grant Funding
- Wellcome Trust
References
This article includes 50 references
- Walker S, Greaves R, O'Hare P. Transcriptional activation by the acidic domain of Vmw65 requires the integrity of the domain and involves additional determinants distinct from those necessary for TFIIB binding.. Mol Cell Biol 1993 Sep;13(9):5233-44.
- Moriuchi H, Moriuchi M, Straus SE, Cohen JI. Varicella-zoster virus open reading frame 10 protein, the herpes simplex virus VP16 homolog, transactivates herpesvirus immediate-early gene promoters.. J Virol 1993 May;67(5):2739-46.
- Lewis JB, Thompson YG, Caughman GB. Transcriptional control of the equine herpesvirus 1 immediate early gene.. Virology 1993 Dec;197(2):788-92.
- Heine JW, Honess RW, Cassai E, Roizman B. Proteins specified by herpes simplex virus. XII. The virion polypeptides of type 1 strains.. J Virol 1974 Sep;14(3):640-51.
- Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.. Mol Cell Biol 1982 Sep;2(9):1044-51.
- Kristie TM, Roizman B. Separation of sequences defining basal expression from those conferring alpha gene recognition within the regulatory domains of herpes simplex virus 1 alpha genes.. Proc Natl Acad Sci U S A 1984 Jul;81(13):4065-9.
- Campbell ME, Palfreyman JW, Preston CM. Identification of herpes simplex virus DNA sequences which encode a trans-acting polypeptide responsible for stimulation of immediate early transcription.. J Mol Biol 1984 Nov 25;180(1):1-19.
- Pellett PE, McKnight JL, Jenkins FJ, Roizman B. Nucleotide sequence and predicted amino acid sequence of a protein encoded in a small herpes simplex virus DNA fragment capable of trans-inducing alpha genes.. Proc Natl Acad Sci U S A 1985 Sep;82(17):5870-4.
- Davison AJ, Scott JE. The complete DNA sequence of varicella-zoster virus.. J Gen Virol 1986 Sep;67 ( Pt 9):1759-816.
- Staudt LM, Singh H, Sen R, Wirth T, Sharp PA, Baltimore D. A lymphoid-specific protein binding to the octamer motif of immunoglobulin genes.. Nature 1986 Oct 16-22;323(6089):640-3.
- Kristie TM, Roizman B. Host cell proteins bind to the cis-acting site required for virion-mediated induction of herpes simplex virus 1 alpha genes.. Proc Natl Acad Sci U S A 1987 Jan;84(1):71-5.
- Gray WL, Baumann RP, Robertson AT, Caughman GB, O'Callaghan DJ, Staczek J. Regulation of equine herpesvirus type 1 gene expression: characterization of immediate early, early, and late transcription.. Virology 1987 May;158(1):79-87.
- Cullen BR. Use of eukaryotic expression technology in the functional analysis of cloned genes.. Methods Enzymol 1987;152:684-704.
- McKnight JL, Kristie TM, Roizman B. Binding of the virion protein mediating alpha gene induction in herpes simplex virus 1-infected cells to its cis site requires cellular proteins.. Proc Natl Acad Sci U S A 1987 Oct;84(20):7061-5.
- Preston CM, Frame MC, Campbell ME. A complex formed between cell components and an HSV structural polypeptide binds to a viral immediate early gene regulatory DNA sequence.. Cell 1988 Feb 12;52(3):425-34.
- O'Hare P, Goding CR. Herpes simplex virus regulatory elements and the immunoglobulin octamer domain bind a common factor and are both targets for virion transactivation.. Cell 1988 Feb 12;52(3):435-45.
- Adams SE, Johnson ID, Braddock M, Kingsman AJ, Kingsman SM, Edwards RM. Synthesis of a gene for the HIV transactivator protein TAT by a novel single stranded approach involving in vivo gap repair.. Nucleic Acids Res 1988 May 25;16(10):4287-98.
- McGeoch DJ, Dalrymple MA, Davison AJ, Dolan A, Frame MC, McNab D, Perry LJ, Scott JE, Taylor P. The complete DNA sequence of the long unique region in the genome of herpes simplex virus type 1.. J Gen Virol 1988 Jul;69 ( Pt 7):1531-74.
- Gerster T, Roeder RG. A herpesvirus trans-activating protein interacts with transcription factor OTF-1 and other cellular proteins.. Proc Natl Acad Sci U S A 1988 Sep;85(17):6347-51.
- Triezenberg SJ, Kingsbury RC, McKnight SL. Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression.. Genes Dev 1988 Jun;2(6):718-29.
- Triezenberg SJ, LaMarco KL, McKnight SL. Evidence of DNA: protein interactions that mediate HSV-1 immediate early gene activation by VP16.. Genes Dev 1988 Jun;2(6):730-42.
- Purewal AS, Allsopp R, Riggio M, Telford EA, Azam S, Davison AJ, Edington N. Equid herpesviruses 1 and 4 encode functional homologs of the herpes simplex virus type 1 virion transactivator protein, VP16.. Virology 1994 Jan;198(1):385-9.
- Friedman AD, Triezenberg SJ, McKnight SL. Expression of a truncated viral trans-activator selectively impedes lytic infection by its cognate virus.. Nature 1988 Sep 29;335(6189):452-4.
- Sadowski I, Ma J, Triezenberg S, Ptashne M. GAL4-VP16 is an unusually potent transcriptional activator.. Nature 1988 Oct 6;335(6190):563-4.
- Ace CI, Dalrymple MA, Ramsay FH, Preston VG, Preston CM. Mutational analysis of the herpes simplex virus type 1 trans-inducing factor Vmw65.. J Gen Virol 1988 Oct;69 ( Pt 10):2595-605.
- Herr W, Sturm RA, Clerc RG, Corcoran LM, Baltimore D, Sharp PA, Ingraham HA, Rosenfeld MG, Finney M, Ruvkun G. The POU domain: a large conserved region in the mammalian pit-1, oct-1, oct-2, and Caenorhabditis elegans unc-86 gene products.. Genes Dev 1988 Dec;2(12A):1513-6.
- Greaves R, O'Hare P. Separation of requirements for protein-DNA complex assembly from those for functional activity in the herpes simplex virus regulatory protein Vmw65.. J Virol 1989 Apr;63(4):1641-50.
- Ace CI, McKee TA, Ryan JM, Cameron JM, Preston CM. Construction and characterization of a herpes simplex virus type 1 mutant unable to transinduce immediate-early gene expression.. J Virol 1989 May;63(5):2260-9.
- O'Hare P, Goding CR, Haigh A. Direct combinatorial interaction between a herpes simplex virus regulatory protein and a cellular octamer-binding factor mediates specific induction of virus immediate-early gene expression.. EMBO J 1988 Dec 20;7(13):4231-8.
- Werstuck G, Capone JP. Mutational analysis of the herpes simplex virus trans-inducing factor Vmw65.. Gene 1989 Feb 20;75(2):213-24.
- Cousens DJ, Greaves R, Goding CR, O'Hare P. The C-terminal 79 amino acids of the herpes simplex virus regulatory protein, Vmw65, efficiently activate transcription in yeast and mammalian cells in chimeric DNA-binding proteins.. EMBO J 1989 Aug;8(8):2337-42.
- Kristie TM, LeBowitz JH, Sharp PA. The octamer-binding proteins form multi-protein--DNA complexes with the HSV alpha TIF regulatory protein.. EMBO J 1989 Dec 20;8(13):4229-38.
- McKee TA, Disney GH, Everett RD, Preston CM. Control of expression of the varicella-zoster virus major immediate early gene.. J Gen Virol 1990 Apr;71 ( Pt 4):897-906.
- Stringer KF, Ingles CJ, Greenblatt J. Direct and selective binding of an acidic transcriptional activation domain to the TATA-box factor TFIID.. Nature 1990 Jun 28;345(6278):783-6.
- Xiao P, Capone JP. A cellular factor binds to the herpes simplex virus type 1 transactivator Vmw65 and is required for Vmw65-dependent protein-DNA complex assembly with Oct-1.. Mol Cell Biol 1990 Sep;10(9):4974-7.
- Cress WD, Triezenberg SJ. Critical structural elements of the VP16 transcriptional activation domain.. Science 1991 Jan 4;251(4989):87-90.
- Kristie TM, Sharp PA. Interactions of the Oct-1 POU subdomains with specific DNA sequences and with the HSV alpha-trans-activator protein.. Genes Dev 1990 Dec;4(12B):2383-96.
- Andrews NC, Faller DV. A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells.. Nucleic Acids Res 1991 May 11;19(9):2499.
- Ingles CJ, Shales M, Cress WD, Triezenberg SJ, Greenblatt J. Reduced binding of TFIID to transcriptionally compromised mutants of VP16.. Nature 1991 Jun 13;351(6327):588-90.
- Lin YS, Ha I, Maldonado E, Reinberg D, Green MR. Binding of general transcription factor TFIIB to an acidic activating region.. Nature 1991 Oct 10;353(6344):569-71.
- Elton DM, Bonass WA, Killington RA, Meredith DM, Halliburton IW. Location of open reading frames coding for equine herpesvirus type-1 glycoproteins with homology to gE and gI of herpes simplex virus.. Am J Vet Res 1991 Aug;52(8):1252-7.
- Weinheimer SP, Boyd BA, Durham SK, Resnick JL, O'Boyle DR 2nd. Deletion of the VP16 open reading frame of herpes simplex virus type 1.. J Virol 1992 Jan;66(1):258-69.
- Kinchington PR, Hougland JK, Arvin AM, Ruyechan WT, Hay J. The varicella-zoster virus immediate-early protein IE62 is a major component of virus particles.. J Virol 1992 Jan;66(1):359-66.
- Purewal AS, Smallwood AV, Kaushal A, Adegboye D, Edington N. Identification and control of the cis-acting elements of the immediate early gene of equid herpesvirus type 1.. J Gen Virol 1992 Mar;73 ( Pt 3):513-9.
- Elliott GD, Meredith DM. The herpes simplex virus type 1 tegument protein VP22 is encoded by gene UL49.. J Gen Virol 1992 Mar;73 ( Pt 3):723-6.
- O'Hare P, Williams G. Structural studies of the acidic transactivation domain of the Vmw65 protein of herpes simplex virus using 1H NMR.. Biochemistry 1992 Apr 28;31(16):4150-6.
- Telford EA, Watson MS, McBride K, Davison AJ. The DNA sequence of equine herpesvirus-1.. Virology 1992 Jul;189(1):304-16.
- Hayes S, O'Hare P. Mapping of a major surface-exposed site in herpes simplex virus protein Vmw65 to a region of direct interaction in a transcription complex assembly.. J Virol 1993 Feb;67(2):852-62.
- Regier JL, Shen F, Triezenberg SJ. Pattern of aromatic and hydrophobic amino acids critical for one of two subdomains of the VP16 transcriptional activator.. Proc Natl Acad Sci U S A 1993 Feb 1;90(3):883-7.
- Goodrich JA, Hoey T, Thut CJ, Admon A, Tjian R. Drosophila TAFII40 interacts with both a VP16 activation domain and the basal transcription factor TFIIB.. Cell 1993 Nov 5;75(3):519-30.
Citations
This article has been cited 15 times.- Feng Z, Zhou F, Tan M, Wang T, Chen Y, Xu W, Li B, Wang X, Deng X, He ML. Targeting m6A modification inhibits herpes virus 1 infection. Genes Dis 2022 Jul;9(4):1114-1128.
- Kim S, Dai G, O'Callaghan DJ, Kim SK. Characterization of cis-acting elements required for autorepression of the equine herpesvirus 1 IE gene. Virus Res 2012 Apr;165(1):52-60.
- Ahn BC, Zhang Y, O'Callaghan DJ. The equine herpesvirus-1 (EHV-1) IR3 transcript downregulates expression of the IE gene and the absence of IR3 gene expression alters EHV-1 biological properties and virulence. Virology 2010 Jul 5;402(2):327-37.
- Ahn BC, Breitenbach JE, Kim SK, O'Callaghan DJ. The equine herpesvirus-1 IR3 gene that lies antisense to the sole immediate-early (IE) gene is trans-activated by the IE protein, and is poorly expressed to a protein. Virology 2007 Jun 20;363(1):15-25.
- Tyack SG, Studdert MJ, Johnson MA. Sequence and function of canine herpesvirus alpha-transinducing factor and its interaction with an immediate early promoter. Virus Genes 2006 Dec;33(3):299-307.
- von Einem J, Schumacher D, O'Callaghan DJ, Osterrieder N. The alpha-TIF (VP16) homologue (ETIF) of equine herpesvirus 1 is essential for secondary envelopment and virus egress. J Virol 2006 Mar;80(6):2609-20.
- Boyer JL, Swaminathan S, Silverstein SJ. The Epstein-Barr virus SM protein is functionally similar to ICP27 from herpes simplex virus in viral infections. J Virol 2002 Sep;76(18):9420-33.
- Gillette K, Misra V, Bratanich A. Sequence analysis of the alpha trans-inducing factor of bovine herpesvirus type 5 (BHV-5). Virus Genes 2002 Mar;24(2):149-52.
- Grapes M, O'Hare P. Differences in determinants required for complex formation and transactivation in related VP16 proteins. J Virol 2000 Nov;74(21):10112-21.
- Huang CC, Herr W. Differential control of transcription by homologous homeodomain coregulators. Mol Cell Biol 1996 Jun;16(6):2967-76.
- Smith RH, Holden VR, O'Callaghan DJ. Nuclear localization and transcriptional activation activities of truncated versions of the immediate-early gene product of equine herpesvirus 1. J Virol 1995 Jun;69(6):3857-62.
- Misra V, Walker S, Hayes S, O'Hare P. The bovine herpesvirus alpha gene trans-inducing factor activates transcription by mechanisms different from those of its herpes simplex virus type 1 counterpart VP16. J Virol 1995 Sep;69(9):5209-16.
- Moriuchi H, Moriuchi M, Pichyangkura R, Triezenberg SJ, Straus SE, Cohen JI. Hydrophobic cluster analysis predicts an amino-terminal domain of varicella-zoster virus open reading frame 10 required for transcriptional activation. Proc Natl Acad Sci U S A 1995 Sep 26;92(20):9333-7.
- Elliott G, Mouzakitis G, O'Hare P. VP16 interacts via its activation domain with VP22, a tegument protein of herpes simplex virus, and is relocated to a novel macromolecular assembly in coexpressing cells. J Virol 1995 Dec;69(12):7932-41.
- Maeda M, Abe M, Aoshima K, Kobayashi A, Fukushi H, Kimura T. Identification of the Promoter Antisense Transcript Enhancing the Transcription of the Equine Herpesvirus-1 Immediate-Early Gene. Viruses 2024 Jul 25;16(8).
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists