1887

Abstract

RNA-remodelling proteins, including RNA helicases and chaperones, function to remodel structured RNAs and/or RNA–protein interactions and play indispensable roles in viral life cycles. Guaico Culex virus (GCXV) is the first uncovered animal-infected multicomponent virus with segmented positive-sense genomic RNAs. GCXV belongs to the Jingmenvirus group, a diverse clade of segmented viruses that are related to the prototypically unsegmented . However, little is known about the exact functions of the GCXV-encoded proteins. Here, we show that the putative non-structural protein (NSP) 2 on segment 2 of GCXV functions as an RNA helicase that unwinds RNA helix bidirectionally in an adenosine triphosphate (ATP)-dependent manner, and an RNA chaperone that remodels structured RNAs and facilitates RNA strand annealing independently of ATP. Together, our findings are the first demonstration of RNA-remodelling activity encoded by Jingmenvirus and highlight the functional significance of NSP2 in the GCXV life cycle.

Funding
This study was supported by the:
  • Youth Innovation Promotion Association of the Chinese Academy of Sciences (Award 2020332)
    • Principle Award Recipient: YangQiu
  • National Natural Science Foundation of China (Award 31970169)
    • Principle Award Recipient: xizhou
  • National Natural Science Foundation of China (Award 81873964)
    • Principle Award Recipient: YangQiu
  • Strategic Priority Research Program of CAS (Award XDB29010300)
    • Principle Award Recipient: xizhou
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001589
2021-04-23
2024-03-28
Loading full text...

Full text loading...

References

  1. Fulton RW. Biological significance of multicomponent viruses. Annu Rev Phytopathol 1980; 18:131–146 [View Article]
    [Google Scholar]
  2. Qin X-C, Shi M, Tian J-H, Lin X-D, Gao D-Y et al. A tick-borne segmented RNA virus contains genome segments derived from unsegmented viral ancestors. Proc Natl Acad Sci U S A 2014; 111:6744–6749 [View Article][PubMed]
    [Google Scholar]
  3. Shi M, Lin X-D, Vasilakis N, Tian J-H, Li C-X et al. Divergent viruses discovered in arthropods and vertebrates revise the evolutionary history of the Flaviviridae and related viruses. J Virol 2016; 90:659–669 [View Article][PubMed]
    [Google Scholar]
  4. Ladner JT, Wiley MR, Beitzel B, Auguste AJ, Dupuis AP et al. A multicomponent animal virus isolated from mosquitoes. Cell Host Microbe 2016; 20:357–367 [View Article][PubMed]
    [Google Scholar]
  5. Falgout B, Pethel M, Zhang YM, Lai CJ. Both nonstructural proteins NS2B and NS3 are required for the proteolytic processing of dengue virus nonstructural proteins. J Virol 1991; 65:2467–2475 [View Article][PubMed]
    [Google Scholar]
  6. Assenberg R, Mastrangelo E, Walter TS, Verma A, Milani M et al. Crystal structure of a novel conformational state of the flavivirus NS3 protein: implications for polyprotein processing and viral replication. J Virol 2009; 83:12895–12906 [View Article][PubMed]
    [Google Scholar]
  7. Wengler G, Czaya G, Färber PM, Hegemann JH. In vitro synthesis of West Nile virus proteins indicates that the amino-terminal segment of the NS3 protein contains the active centre of the protease which cleaves the viral polyprotein after multiple basic amino acids. J Gen Virol 1991; 72:851–858 [View Article][PubMed]
    [Google Scholar]
  8. Chambers TJ, Nestorowicz A, Amberg SM, Rice CM. Mutagenesis of the yellow fever virus NS2B protein: effects on proteolytic processing, NS2B-NS3 complex formation, and viral replication. J Virol 1993; 67:6797–6807 [View Article][PubMed]
    [Google Scholar]
  9. Gorbalenya AE, Donchenko AP, Koonin EV, Blinov VM. N-terminal domains of putative helicases of flavi- and pestiviruses may be serine proteases. Nucleic Acids Res 1989; 17:3889–3897 [View Article][PubMed]
    [Google Scholar]
  10. Bleichert F, Baserga SJ. The long unwinding road of RNA helicases. Mol Cell 2007; 27:339–352 [View Article][PubMed]
    [Google Scholar]
  11. Jarmoskaite I, Russell R. RNA helicase proteins as chaperones and remodelers. Annu Rev Biochem 2014; 83:697–725 [View Article][PubMed]
    [Google Scholar]
  12. Grohman JK, Gorelick RJ, Lickwar CR, Lieb JD, Bower BD et al. A guanosine-centric mechanism for RNA chaperone function. Science 2013; 340:190–195 [View Article][PubMed]
    [Google Scholar]
  13. Lorsch JR. RNA chaperones exist and dead box proteins get a life. Cell 2002; 109:797–800 [View Article][PubMed]
    [Google Scholar]
  14. Rajkowitsch L, Chen D, Stampfl S, Semrad K, Waldsich C et al. Rna chaperones, RNA annealers and RNA helicases. RNA Biol 2007; 4:118–130 [View Article][PubMed]
    [Google Scholar]
  15. Kadaré G, Haenni AL. Virus-encoded RNA helicases. J Virol 1997; 71:2583–2590 [View Article][PubMed]
    [Google Scholar]
  16. Musier-Forsyth K. RNA remodeling by chaperones and helicases. RNA Biol 2010; 7:632–633 [View Article][PubMed]
    [Google Scholar]
  17. Xia H, Wang P, Wang G-C, Yang J, Sun X et al. Human enterovirus nonstructural protein 2CATPase functions as both an RNA helicase and ATP-independent RNA chaperone. PLoS Pathog 2015; 11:e1005067 [View Article][PubMed]
    [Google Scholar]
  18. Matusan AE, Pryor MJ, Davidson AD, Wright PJ. Mutagenesis of the dengue virus type 2 NS3 protein within and outside helicase motifs: effects on enzyme activity and virus replication. J Virol 2001; 75:9633–9643 [View Article][PubMed]
    [Google Scholar]
  19. Karpe YA, Aher PP, Lole KS. NTPase and 5'-RNA triphosphatase activities of Chikungunya virus NSP2 protein. PLoS One 2011; 6:e22336 [View Article][PubMed]
    [Google Scholar]
  20. Tanner JA, Watt RM, Chai Y-B, Lu L-Y, Lin MC et al. The severe acute respiratory syndrome (SARS) coronavirus NTPase/helicase belongs to a distinct class of 5' to 3' viral helicases. J Biol Chem 2003; 278:39578–39582 [View Article][PubMed]
    [Google Scholar]
  21. Shu T, Huang M, Wu D, Ren Y, Zhang X et al. SARS-Coronavirus-2 Nsp13 possesses NTPase and RNA helicase activities that can be inhibited by bismuth salts. Virol Sin 2020; 35:321–329 [View Article][PubMed]
    [Google Scholar]
  22. Li T-F, Hosmillo M, Schwanke H, Shu T, Wang Z et al. Human norovirus NS3 has RNA helicase and chaperoning activities. J Virol 2018; 92: 01 03 2018 [View Article][PubMed]
    [Google Scholar]
  23. Shu T, Gan T, Bai P, Wang X, Qian Q et al. Ebola virus VP35 has novel NTPase and helicase-like activities. Nucleic Acids Res 2019; 47:5837–5851 [View Article][PubMed]
    [Google Scholar]
  24. Wang Q, Han Y, Qiu Y, Zhang S, Tang F et al. Identification and characterization of RNA duplex unwinding and ATPase activities of an alphatetravirus superfamily 1 helicase. Virology 2012; 433:440–448 [View Article][PubMed]
    [Google Scholar]
  25. Sloan KE, Bohnsack MT. Unravelling the mechanisms of RNA helicase regulation. Trends Biochem Sci 2018; 43:237–250 [View Article][PubMed]
    [Google Scholar]
  26. Eagles RM, Balmori-Melián E, Beck DL, Gardner RC, Forster RL. Characterization of NTPase, RNA-binding and RNA-helicase activities of the cytoplasmic inclusion protein of tamarillo mosaic potyvirus. Eur J Biochem 1994; 224:677–684 [View Article][PubMed]
    [Google Scholar]
  27. Laín S, Martín MT, Riechmann JL, García JA. Novel catalytic activity associated with positive-strand RNA virus infection: nucleic acid-stimulated ATPase activity of the plum pox potyvirus helicaselike protein. J Virol 1991; 65:1–6 [View Article][PubMed]
    [Google Scholar]
  28. Suzich JA, Tamura JK, Palmer-Hill F, Warrener P, Grakoui A et al. Hepatitis C virus NS3 protein polynucleotide-stimulated nucleoside triphosphatase and comparison with the related pestivirus and flavivirus enzymes. J Virol 1993; 67:6152–6158 [View Article][PubMed]
    [Google Scholar]
  29. Hirling H, Scheffner M, Restle T, Stahl H. RNA helicase activity associated with the human p68 protein. Nature 1989; 339:562–564 [View Article][PubMed]
    [Google Scholar]
  30. Laín S, Riechmann JL, García JA. RNA helicase: a novel activity associated with a protein encoded by a positive strand RNA virus. Nucleic Acids Res 1990; 18:7003–7006 [View Article][PubMed]
    [Google Scholar]
  31. Lee CG, Hurwitz J. A new RNA helicase isolated from HeLa cells that catalytically translocates in the 3' to 5' direction. J Biol Chem 1992; 267:4398–4407 [View Article][PubMed]
    [Google Scholar]
  32. Scheffner M, Knippers R, Stahl H. RNA unwinding activity of SV40 large T antigen. Cell 1989; 57:955–963 [View Article][PubMed]
    [Google Scholar]
  33. Shuman S. Vaccinia virus RNA helicase. directionality and substrate specificity. J Biol Chem 1993; 268:11798–11802 [View Article][PubMed]
    [Google Scholar]
  34. Warrener P, Collett MS. Pestivirus NS3 (p80) protein possesses RNA helicase activity. J Virol 1995; 69:1720–1726 [View Article][PubMed]
    [Google Scholar]
  35. Tai CL, Chi WK, Chen DS, Hwang LH. The helicase activity associated with hepatitis C virus nonstructural protein 3 (NS3). J Virol 1996; 70:8477–8484 [View Article][PubMed]
    [Google Scholar]
  36. Gebhard LG, Kaufman SB, Gamarnik AV. Novel ATP-independent RNA annealing activity of the dengue virus NS3 helicase. PLoS One 2012; 7:e36244 [View Article][PubMed]
    [Google Scholar]
  37. Wang Z-D, Wang B, Wei F, Han S-Z, Zhang L et al. A new segmented virus associated with human febrile illness in China. N Engl J Med 2019; 380:2116–2125 [View Article][PubMed]
    [Google Scholar]
  38. Zhou H, Qian Q, Shu T, Xu J, Kong J et al. Hepatitis C virus NS2 protein suppresses RNA interference in cells. Virol Sin 2020; 35:436–444 [View Article][PubMed]
    [Google Scholar]
  39. Kim SW, Li Z, Moore PS, Monaghan AP, Chang Y et al. A sensitive non-radioactive northern blot method to detect small RNAs. Nucleic Acids Res 2010; 38:e98 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001589
Loading
/content/journal/jgv/10.1099/jgv.0.001589
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error