FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Viruses / Cloning the Horse RNA Polymerase I Promoter and Its Applicat...
Viruses2016; 8(6); 119; doi: 10.3390/v8060119

Cloning the Horse RNA Polymerase I Promoter and Its Application to Studying Influenza Virus Polymerase Activity.

Abstract: An influenza virus polymerase reconstitution assay based on the human, dog, or chicken RNA polymerase I (PolI) promoter has been developed and widely used to study the polymerase activity of the influenza virus in corresponding cell types. Although it is an important member of the influenza virus family and has been known for sixty years, no studies have been performed to clone the horse PolI promoter or to study the polymerase activity of equine influenza virus (EIV) in horse cells. In our study, the horse RNA PolI promoter was cloned from fetal equine lung cells. Using the luciferase assay, it was found that a 500 bp horse RNA PolI promoter sequence was required for efficient transcription. Then, using the developed polymerase reconstitution assay based on the horse RNA PolI promoter, the polymerase activity of two EIV strains was compared, and equine myxovirus resistance A protein was identified as having the inhibiting EIV polymerase activity function in horse cells. Our study enriches our knowledge of the RNA PolI promoter of eukaryotic species and provides a useful tool for the study of influenza virus polymerase activity in horse cells.
Get Full Text
Publication Date: 2016-05-31 PubMed ID: 27258298PubMed Central: PMC4926170DOI: 10.3390/v8060119Google 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.
LinkedInCopy
  • 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.

This research article revolves around the cloning of horse RNA Polymerase I (PolI) promoter and its application in evaluating the polymerase activity of the equine influenza virus in horse cells.

Cloning of the Horse RNA PolI Promoter

  • In this study, the horse RNA PolI promoter was cloned from fetal equine lung cells. This is a significant step as until now, no studies had been conducted to clone the horse PolI promoter.
  • The cloning of the horse RNA PolI promoter expands our understanding of the RNA PolI promoter in eukaryotic species, which are organisms whose cells have a nucleus enclosed within membranes. This includes animals, plants and fungi.

Studying Equine Influenza Virus Polymerase Activity

  • Previously, a reconstitution assay based on the human, dog or chicken RNA PolI promoter has been used to study the polymerase activity of the influenza virus in corresponding cell types.
  • In this research, a similar polymerase reconstitution assay, but based on the newly cloned horse RNA PolI promoter, was developed for studying the polymerase activity of the equine influenza virus (EIV) in horse cells.
  • The relative activity of two EIV strains was compared using this novel assay.

Identifying the Inhibition Function of Equine Myxovirus Resistance A Protein

  • Furthermore, the research identified the protein “equine myxovirus resistance A” as having the ability to inhibit EIV polymerase activity in horse cells.
  • This discovery can provide valuable input in the study of the response of horse cells to EIV and may guide future development of remedies or preventatives against EIV.

Further Implication of the Study

  • This study not only provides deeper insights about RNA PolI promoters among eukaryotes, but it also offers a useful tool for future research on influenza virus polymerase activity in horse cells.
  • This opens new avenues for better understanding and treating influenza viruses, particularly the ones impacting equine species.

Cite This Article

APA
Lu G, He D, Wang Z, Ou S, Yuan R, Li S. (2016). Cloning the Horse RNA Polymerase I Promoter and Its Application to Studying Influenza Virus Polymerase Activity. Viruses, 8(6), 119. https://doi.org/10.3390/v8060119

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 8
Issue: 6
PII: 119

Researcher Affiliations

Lu, Gang
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. 18814113742@163.com.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. 18814113742@163.com.
He, Dong
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. donghe9108@163.com.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. donghe9108@163.com.
Wang, Zengchao
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. lugang19860501@163.com.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. lugang19860501@163.com.
Ou, Shudan
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. whj751364486@163.com.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. whj751364486@163.com.
Yuan, Rong
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. jeremyjessica@163.com.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. jeremyjessica@163.com.
Li, Shoujun
  • College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. shoujunli@scau.edu.cn.
  • Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. shoujunli@scau.edu.cn.

MeSH Terms

  • Animals
  • Cell Line
  • Cloning, Molecular
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • Horses
  • Influenza A virus / enzymology
  • Influenza A virus / physiology
  • Promoter Regions, Genetic
  • RNA Polymerase I / genetics
  • Transcription, Genetic
  • Virus Replication

References

This article includes 35 references
  1. Eisfeld AJ, Neumann G, Kawaoka Y. At the centre: influenza A virus ribonucleoproteins.. Nat Rev Microbiol 2015 Jan;13(1):28-41.
    doi: 10.1038/nrmicro3367pmc: PMC5619696pubmed: 25417656google scholar: lookup
  2. Neumann G, Watanabe T, Ito H, Watanabe S, Goto H, Gao P, Hughes M, Perez DR, Donis R, Hoffmann E, Hobom G, Kawaoka Y. Generation of influenza A viruses entirely from cloned cDNAs.. Proc Natl Acad Sci U S A 1999 Aug 3;96(16):9345-50.
    doi: 10.1073/pnas.96.16.9345pmc: PMC17785pubmed: 10430945google scholar: lookup
  3. Fodor E, Devenish L, Engelhardt OG, Palese P, Brownlee GG, García-Sastre A. Rescue of influenza A virus from recombinant DNA.. J Virol 1999 Nov;73(11):9679-82.
    pmc: PMC113010pubmed: 10516084doi: 10.1128/jvi.73.11.9679-9682.1999google scholar: lookup
  4. Hoffmann E, Neumann G, Kawaoka Y, Hobom G, Webster RG. A DNA transfection system for generation of influenza A virus from eight plasmids.. Proc Natl Acad Sci U S A 2000 May 23;97(11):6108-13.
    doi: 10.1073/pnas.100133697pmc: PMC18566pubmed: 10801978google scholar: lookup
  5. Massin P, Rodrigues P, Marasescu M, van der Werf S, Naffakh N. Cloning of the chicken RNA polymerase I promoter and use for reverse genetics of influenza A viruses in avian cells.. J Virol 2005 Nov;79(21):13811-6.
    doi: 10.1128/JVI.79.21.13811-13816.2005pmc: PMC1262576pubmed: 16227302google scholar: lookup
  6. Murakami S, Horimoto T, Yamada S, Kakugawa S, Goto H, Kawaoka Y. Establishment of canine RNA polymerase I-driven reverse genetics for influenza A virus: its application for H5N1 vaccine production.. J Virol 2008 Feb;82(3):1605-9.
    doi: 10.1128/JVI.01876-07pmc: PMC2224466pubmed: 18045936google scholar: lookup
  7. Wang Z, Duke GM. Cloning of the canine RNA polymerase I promoter and establishment of reverse genetics for influenza A and B in MDCK cells.. Virol J 2007 Oct 23;4:102.
    doi: 10.1186/1743-422X-4-102pmc: PMC2241602pubmed: 17956624google scholar: lookup
  8. Doelling JH, Pikaard CS. Species-specificity of rRNA gene transcription in plants manifested as a switch in RNA polymerase specificity.. Nucleic Acids Res 1996 Dec 1;24(23):4725-32.
    doi: 10.1093/nar/24.23.4725pmc: PMC146310pubmed: 8972859google scholar: lookup
  9. Zhang X, Curtiss R 3rd. Efficient generation of influenza virus with a mouse RNA polymerase I-driven all-in-one plasmid.. Virol J 2015 Jun 22;12:95.
    doi: 10.1186/s12985-015-0321-5pmc: PMC4495709pubmed: 26093583google scholar: lookup
  10. Gabriel G, Dauber B, Wolff T, Planz O, Klenk HD, Stech J. The viral polymerase mediates adaptation of an avian influenza virus to a mammalian host.. Proc Natl Acad Sci U S A 2005 Dec 20;102(51):18590-5.
    doi: 10.1073/pnas.0507415102pmc: PMC1317936pubmed: 16339318google scholar: lookup
  11. König R, Stertz S, Zhou Y, Inoue A, Hoffmann HH, Bhattacharyya S, Alamares JG, Tscherne DM, Ortigoza MB, Liang Y, Gao Q, Andrews SE, Bandyopadhyay S, De Jesus P, Tu BP, Pache L, Shih C, Orth A, Bonamy G, Miraglia L, Ideker T, García-Sastre A, Young JA, Palese P, Shaw ML, Chanda SK. Human host factors required for influenza virus replication.. Nature 2010 Feb 11;463(7282):813-7.
    doi: 10.1038/nature08699pmc: PMC2862546pubmed: 20027183google scholar: lookup
  12. Tong S, Li Y, Rivailler P, Conrardy C, Castillo DA, Chen LM, Recuenco S, Ellison JA, Davis CT, York IA, Turmelle AS, Moran D, Rogers S, Shi M, Tao Y, Weil MR, Tang K, Rowe LA, Sammons S, Xu X, Frace M, Lindblade KA, Cox NJ, Anderson LJ, Rupprecht CE, Donis RO. A distinct lineage of influenza A virus from bats.. Proc Natl Acad Sci U S A 2012 Mar 13;109(11):4269-74.
    doi: 10.1073/pnas.1116200109pmc: PMC3306675pubmed: 22371588google scholar: lookup
  13. Moncorgé O, Mura M, Barclay WS. Evidence for avian and human host cell factors that affect the activity of influenza virus polymerase.. J Virol 2010 Oct;84(19):9978-86.
    doi: 10.1128/JVI.01134-10pmc: PMC2937815pubmed: 20631125google scholar: lookup
  14. Bussey KA, Bousse TL, Desmet EA, Kim B, Takimoto T. PB2 residue 271 plays a key role in enhanced polymerase activity of influenza A viruses in mammalian host cells.. J Virol 2010 May;84(9):4395-406.
    doi: 10.1128/JVI.02642-09pmc: PMC2863787pubmed: 20181719google scholar: lookup
  15. Gabriel G, Abram M, Keiner B, Wagner R, Klenk HD, Stech J. Differential polymerase activity in avian and mammalian cells determines host range of influenza virus.. J Virol 2007 Sep;81(17):9601-4.
    doi: 10.1128/JVI.00666-07pmc: PMC1951401pubmed: 17567688google scholar: lookup
  16. Song J, Feng H, Xu J, Zhao D, Shi J, Li Y, Deng G, Jiang Y, Li X, Zhu P, Guan Y, Bu Z, Kawaoka Y, Chen H. The PA protein directly contributes to the virulence of H5N1 avian influenza viruses in domestic ducks.. J Virol 2011 Mar;85(5):2180-8.
    doi: 10.1128/JVI.01975-10pmc: PMC3067757pubmed: 21177821google scholar: lookup
  17. Lu G, Guo W, Qi T, Ma J, Zhao S, Tian Z, Pan J, Zhu C, Wang X, Xiang W. Genetic analysis of the PB1-F2 gene of equine influenza virus.. Virus Genes 2013 Oct;47(2):250-8.
    doi: 10.1007/s11262-013-0935-xpubmed: 23780220google scholar: lookup
  18. Yin X, Lu G, Guo W, Qi T, Ma J, Zhu C, Zhao S, Pan J, Xiang W. Identification of equine influenza virus infection in Asian wild horses (Equus przewalskii).. Arch Virol 2014 May;159(5):1159-62.
    doi: 10.1007/s00705-013-1908-zpubmed: 24212887google scholar: lookup
  19. van Maanen C, Cullinane A. Equine influenza virus infections: an update.. Vet Q 2002 Jun;24(2):79-94.
    doi: 10.1080/01652176.2002.9695127pubmed: 12095083google scholar: lookup
  20. Lai AC, Chambers TM, Holland RE Jr, Morley PS, Haines DM, Townsend HG, Barrandeguy M. Diverged evolution of recent equine-2 influenza (H3N8) viruses in the Western Hemisphere.. Arch Virol 2001;146(6):1063-74.
    doi: 10.1007/s007050170106pubmed: 11504416google scholar: lookup
  21. Grummt I. Life on a planet of its own: regulation of RNA polymerase I transcription in the nucleolus.. Genes Dev 2003 Jul 15;17(14):1691-702.
    doi: 10.1101/gad.1098503Rpubmed: 12865296google scholar: lookup
  22. Bilofsky HS, Burks C. The GenBank genetic sequence data bank.. Nucleic Acids Res 1988 Mar 11;16(5):1861-3.
    doi: 10.1093/nar/16.5.1861pmc: PMC338181pubmed: 3353225google scholar: lookup
  23. Hubbard T, Barker D, Birney E, Cameron G, Chen Y, Clark L, Cox T, Cuff J, Curwen V, Down T, Durbin R, Eyras E, Gilbert J, Hammond M, Huminiecki L, Kasprzyk A, Lehvaslaiho H, Lijnzaad P, Melsopp C, Mongin E, Pettett R, Pocock M, Potter S, Rust A, Schmidt E, Searle S, Slater G, Smith J, Spooner W, Stabenau A, Stalker J, Stupka E, Ureta-Vidal A, Vastrik I, Clamp M. The Ensembl genome database project.. Nucleic Acids Res 2002 Jan 1;30(1):38-41.
    doi: 10.1093/nar/30.1.38pmc: PMC99161pubmed: 11752248google scholar: lookup
  24. Hall T. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 1999;41:95–98.
  25. Crooks GE, Hon G, Chandonia JM, Brenner SE. WebLogo: a sequence logo generator.. Genome Res 2004 Jun;14(6):1188-90.
    doi: 10.1101/gr.849004pmc: PMC419797pubmed: 15173120google scholar: lookup
  26. KITCHEN RH, KEHLER WH, HENTHORNE JC. THE 1963 EQUINE INFLUENZA EPIZOOTIC.. J Am Vet Med Assoc 1963 Nov 15;143:1108-10.
    pubmed: 14075417
  27. Prism, GraphPad. GraphPad Software Inc.; San Diego, CA, USA: 2007. version 5.01.
  28. Su S, Wang L, Fu X, He S, Hong M, Zhou P, Lai A, Gray G, Li S. Equine influenza A(H3N8) virus infection in cats.. Emerg Infect Dis 2014 Dec;20(12):2096-9.
    doi: 10.3201/eid2012.140867pmc: PMC4257791pubmed: 25417790google scholar: lookup
  29. Xiao H, Killip MJ, Staeheli P, Randall RE, Jackson D. The human interferon-induced MxA protein inhibits early stages of influenza A virus infection by retaining the incoming viral genome in the cytoplasm.. J Virol 2013 Dec;87(23):13053-8.
    doi: 10.1128/JVI.02220-13pmc: PMC3838145pubmed: 24049170google scholar: lookup
  30. Haller O, Stertz S, Kochs G. The Mx GTPase family of interferon-induced antiviral proteins.. Microbes Infect 2007 Nov-Dec;9(14-15):1636-43.
    doi: 10.1016/j.micinf.2007.09.010pubmed: 18062906google scholar: lookup
  31. Ko JH, Takada A, Mitsuhashi T, Agui T, Watanabe T. Native antiviral specificity of chicken Mx protein depends on amino acid variation at position 631.. Anim Genet 2004 Apr;35(2):119-22.
    doi: 10.1111/j.1365-2052.2004.01096.xpubmed: 15025571google scholar: lookup
  32. Verhelst J, Parthoens E, Schepens B, Fiers W, Saelens X. Interferon-inducible protein Mx1 inhibits influenza virus by interfering with functional viral ribonucleoprotein complex assembly.. J Virol 2012 Dec;86(24):13445-55.
    doi: 10.1128/JVI.01682-12pmc: PMC3503048pubmed: 23015724google scholar: lookup
  33. Miotto O, Heiny AT, Albrecht R, García-Sastre A, Tan TW, August JT, Brusic V. Complete-proteome mapping of human influenza A adaptive mutations: implications for human transmissibility of zoonotic strains.. PLoS One 2010 Feb 3;5(2):e9025.
    doi: 10.1371/journal.pone.0009025pmc: PMC2815782pubmed: 20140252google scholar: lookup
  34. Ma J, Liu K, Xue C, Zhou J, Xu S, Ren Y, Zheng J, Cao Y. Impact of the segment-specific region of the 3'-untranslated region of the influenza A virus PB1 segment on protein expression.. Virus Genes 2013 Dec;47(3):429-38.
    doi: 10.1007/s11262-013-0969-0pubmed: 23949786google scholar: lookup
  35. Wang L, Lee CW. Sequencing and mutational analysis of the non-coding regions of influenza A virus.. Vet Microbiol 2009 Mar 30;135(3-4):239-47.
    doi: 10.1016/j.vetmic.2008.09.067pubmed: 18986781google scholar: lookup

Citations

This article has been cited 7 times.
  1. Chiem K, Nogales A, Martinez-Sobrido L. Generation, Characterization, and Applications of Influenza A Reporter Viruses. Methods Mol Biol 2022;2524:249-268.
    doi: 10.1007/978-1-0716-2453-1_19pubmed: 35821477google scholar: lookup
  2. Lu G, Zheng F, Ou J, Yin X, Li S. Investigating Influenza Virus Polymerase Activity in Feline Cells Based on the Influenza Virus Minigenome Replication System Driven by the Feline RNA Polymerase I Promoter. Front Immunol 2022;13:827681.
    doi: 10.3389/fimmu.2022.827681pubmed: 35693765google scholar: lookup
  3. Neumann G. Influenza Reverse Genetics-Historical Perspective. Cold Spring Harb Perspect Med 2021 Apr 1;11(4).
    doi: 10.1101/cshperspect.a038547pubmed: 31964649google scholar: lookup
  4. Wang K, Huang Q, Yang Z, Qi K, Liu H, Chen H. Alternative reverse genetics system for influenza viruses based on a synthesized swine 45S rRNA promoter. Virus Genes 2017 Aug;53(4):661-666.
    doi: 10.1007/s11262-017-1457-8pubmed: 28434065google scholar: lookup
  5. Nogales A, Martínez-Sobrido L. Reverse Genetics Approaches for the Development of Influenza Vaccines. Int J Mol Sci 2016 Dec 22;18(1).
    doi: 10.3390/ijms18010020pubmed: 28025504google scholar: lookup
  6. Jang Y, Baek YG, Lee YN, Cha RM, Choi YC, Park MJ, Lee YJ, Lee EK. Research Note: Establishment of vector system harboring duck RNA polymerase I promoter for avian influenza virus. Poult Sci 2025 Jan;104(1):104570.
    doi: 10.1016/j.psj.2024.104570pubmed: 39631283google scholar: lookup
  7. Zhang X, Yu H, Sun P, Huang M, Li B. Antiviral Effects and Mechanisms of Active Ingredients in Tea. Molecules 2024 Nov 4;29(21).
    doi: 10.3390/molecules29215218pubmed: 39519859google scholar: lookup
Subscribe to our newsletter
Join 99,109 horse owners like you who receive our email updates on horse health and nutrition.