Home LiteratureArticle Details
PMID: 14963133 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Alterations of pr-M cleavage and virus export in pr-M junction chimeric dengue viruses.

Journal of virology ·Vol. 78 ·No. 5 ·2004-03-00 ·Pages 2367-81

Keelapang P, Sriburi R, Supasa S, Panyadee N, Songjaeng A, Jairungsri A, Puttikhunt C, Kasinrerk W, Malasit P, Sittisombut N

Abstract

During the export of flavivirus particles through the secretory pathway, a viral envelope glycoprotein, prM, is cleaved by the proprotein convertase furin; this cleavage is required for the subsequent rearrangement of receptor-binding E glycoprotein and for virus infectivity. Similar to many furin substrates, prM in vector-borne flaviviruses contains basic residues at positions P1, P2, and P4 proximal to the cleavage site; in addition, a number of charged residues are found at position P3 and between positions P5 and P13 that are conserved for each flavivirus antigenic complex. The influence of additional charged residues on pr-M cleavage and virus replication was investigated by replacing the 13-amino-acid, cleavage-proximal region of a dengue virus (strain 16681) with those of tick-borne encephalitis virus (TBEV), yellow fever virus (YFV), and Japanese encephalitis virus (JEV) and by comparing the resultant chimeric viruses generated from RNA-transfected mosquito cells. Among the three chimeric viruses, cleavage of prM was enhanced to a larger extent in JEVpr/16681 than in YFVpr/16681 but was slightly reduced in TBEVpr/16681. Unexpectedly, JEVpr/16681 exhibited decreased focus size, reduced peak titer, and depressed replication in C6/36, PS, and Vero cell lines. The reduction of JEVpr/16681 multiplication correlated with delayed export of infectious virions out of infected cells but not with changes in specific infectivity. Binding of JEVpr/16681 to immobilized heparin and the heparin-inhibitable infection of cells were not altered. Thus, diverse pr-M junction-proximal sequences of flaviviruses differentially influence pr-M cleavage when tested in a dengue virus prM background. More importantly, greatly enhanced prM cleavability adversely affects dengue virus export while exerting a minimal effect on infectivity. Because extensive changes of charged residues at the pr-M junction, as in JEVpr/16681, were not observed among a large number of dengue virus isolates, these results provide a possible mechanism by which the sequence conservation of the pr-M junction of dengue virus is maintained in nature.

MeSH Terms
Amino Acid Sequence Animals Biological Transport Cell Line Chlorocebus aethiops Culicidae/virology DNA, Recombinant/genetics Dengue Virus/drug effects,genetics,physiology Genetic Engineering Heparin/metabolism,pharmacology Molecular Sequence Data Movement Protein Processing, Post-Translational Recombinant Fusion Proteins/chemistry,genetics,metabolism Vero Cells Viral Proteins/chemistry,genetics,metabolism Virus Replication
Chemicals
DNA, Recombinant Recombinant Fusion Proteins Viral Proteins Heparin
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Keelapang Poonsook
Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Bangkok 10400, USA.
Sriburi Roongtawan
Supasa Sanpaechuda
Panyadee Nantaya
Songjaeng Adisak
Jairungsri Aroonroong
Puttikhunt Chunya
Kasinrerk Watchara
Malasit Prida
Sittisombut Nopporn
References (80)
80 references, click to expand
  1. Identification of an epitope on the dengue virus membrane (M) protein defined by cross-protective monoclonal antibodies: design of an improved epitope sequence based on common determinants present in both envelope (E and M) proteins.
    Arch Virol. 1999;144(12):2313-30 PMID: 10664386
  2. Processing, stability, and receptor binding properties of oligomeric envelope glycoprotein from a primary HIV-1 isolate.
    J Biol Chem. 2000 Nov 10;275(45):35137-45 PMID: 10926931
  3. Pandemic threat posed by avian influenza A viruses.
    Clin Microbiol Rev. 2001 Jan;14(1):129-49 PMID: 11148006
  4. Implication of the proprotein convertases furin, PC5 and PC7 in the cleavage of surface glycoproteins of Hong Kong, Ebola and respiratory syncytial viruses: a comparative analysis with fluorogenic peptides.
    Biochem J. 2001 Feb 1;353(Pt 3):537-45 PMID: 11171050
  5. Construction of infectious dengue 2 virus cDNA clones using high copy number plasmid.
    J Virol Methods. 2001 Mar;92(1):71-82 PMID: 11164920
  6. Molecular organization of a recombinant subviral particle from tick-borne encephalitis virus.
    Mol Cell. 2001 Mar;7(3):593-602 PMID: 11463384
  7. Enhancing the proteolytic maturation of human immunodeficiency virus type 1 envelope glycoproteins.
    J Virol. 2002 Mar;76(6):2606-16 PMID: 11861826
  8. Structural investigation of the HIV-1 envelope glycoprotein gp160 cleavage site.
    Chemistry. 2002 Mar 15;8(6):1467-73 PMID: 11921231
  9. Folding and dimerization of tick-borne encephalitis virus envelope proteins prM and E in the endoplasmic reticulum.
    J Virol. 2002 Jun;76(11):5480-91 PMID: 11991976
  10. Furin at the cutting edge: from protein traffic to embryogenesis and disease.
    Nat Rev Mol Cell Biol. 2002 Oct;3(10):753-66 PMID: 12360192
  11. Cleavage of protein prM is necessary for infection of BHK-21 cells by tick-borne encephalitis virus.
    J Gen Virol. 2003 Jan;84(Pt 1):183-91 PMID: 12533715
  12. Structures of immature flavivirus particles.
    EMBO J. 2003 Jun 2;22(11):2604-13 PMID: 12773377
  13. Assessment and application of a cell line from pig kidney for plaque assay and neutralization tests with twelve group B arboviruses.
    Am J Epidemiol. 1966 Nov;84(3):439-56 PMID: 6005905
  14. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  15. Dengue virus-specific and flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence.
    Am J Trop Med Hyg. 1982 Jul;31(4):830-6 PMID: 6285749
  16. Topographical analysis of antigenic determinants on envelope glycoprotein V3 (E) of Japanese encephalitis virus, using monoclonal antibodies.
    J Virol. 1983 Jan;45(1):124-32 PMID: 6185694
  17. Maturation of the dengue-2 virus NS1 protein in insect cells: effects of downstream NS2A sequences on baculovirus-expressed gene constructs.
    J Gen Virol. 1995 Apr;76 ( Pt 4):979-84 PMID: 9049346
  18. Construction of infectious cDNA clones for dengue 2 virus: strain 16681 and its attenuated vaccine derivative, strain PDK-53.
    Virology. 1997 Apr 14;230(2):300-8 PMID: 9143286
  19. Activation of endothelial cells via antibody-enhanced dengue virus infection of peripheral blood monocytes.
    J Virol. 1997 Jun;71(6):4226-32 PMID: 9151809
  20. Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate.
    Nat Med. 1997 Aug;3(8):866-71 PMID: 9256277
  21. Proteolytic activation of tick-borne encephalitis virus by furin.
    J Virol. 1997 Nov;71(11):8475-81 PMID: 9343204
  22. Phylogeny of the genus Flavivirus.
    J Virol. 1998 Jan;72(1):73-83 PMID: 9420202
  23. Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness.
    Science. 1998 Jan 16;279(5349):393-6 PMID: 9430591
  24. Retroviral envelope glycoprotein processing: structural investigation of the cleavage site.
    Biochemistry. 1998 Mar 31;37(13):4510-7 PMID: 9521771
  25. Furin: a mammalian subtilisin/Kex2p-like endoprotease involved in processing of a wide variety of precursor proteins.
    Biochem J. 1997 Nov 1;327 ( Pt 3):625-35 PMID: 9599222
  26. Encapsidation of the flavivirus kunjin replicon RNA by using a complementation system providing Kunjin virus structural proteins in trans.
    J Virol. 1998 Jul;72(7):5967-77 PMID: 9621059
  27. Monoclonal antibody mapping of the envelope glycoprotein of the dengue 2 virus, Jamaica.
    Virology. 1998 Jul 5;246(2):317-28 PMID: 9657950
  28. The proprotein convertases.
    Curr Opin Chem Biol. 1998 Feb;2(1):31-9 PMID: 9667917
  29. Structure of the hemagglutinin precursor cleavage site, a determinant of influenza pathogenicity and the origin of the labile conformation.
    Cell. 1998 Oct 30;95(3):409-17 PMID: 9814710
  30. Structure of dengue virus: implications for flavivirus organization, maturation, and fusion.
    Cell. 2002 Mar 8;108(5):717-25 PMID: 11893341
  31. Immune response to synthetic peptides of dengue prM protein.
    Vaccine. 2002 Mar 15;20(13-14):1823-30 PMID: 11906771
  32. Mutational analysis of a neutralization epitope on the dengue type 2 virus (DEN2) envelope protein: monoclonal antibody resistant DEN2/DEN4 chimeras exhibit reduced mouse neurovirulence.
    Virology. 1996 Oct 15;224(2):437-45 PMID: 8874504
  33. Dengue virus infection of human endothelial cells leads to chemokine production, complement activation, and apoptosis.
    J Immunol. 1998 Dec 1;161(11):6338-46 PMID: 9834124
  34. PrM- and cell-binding domains of the dengue virus E protein.
    J Virol. 1999 Mar;73(3):2547-51 PMID: 9971841
  35. Bi-cycling the furin pathway: from TGN localization to pathogen activation and embryogenesis.
    Trends Cell Biol. 1999 Jan;9(1):28-35 PMID: 10087614
  36. Titration of dengue viruses by immunofluorescence in microtiter plates.
    J Clin Microbiol. 1984 Nov;20(5):1017-9 PMID: 6392318
  37. Epitopic analysis of antigenic determinants on the surface of dengue-2 virions using monoclonal antibodies.
    Am J Trop Med Hyg. 1985 Jan;34(1):162-9 PMID: 2578750
  38. Sequence of simian immunodeficiency virus from macaque and its relationship to other human and simian retroviruses.
    Nature. 1987 Aug 6-12;328(6130):543-7 PMID: 3649576
  39. Evidence that the mature form of the flavivirus nonstructural protein NS1 is a dimer.
    Virology. 1988 Jan;162(1):187-96 PMID: 2827377
  40. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
    Anal Biochem. 1987 Nov 1;166(2):368-79 PMID: 2449095
  41. Cell-associated West Nile flavivirus is covered with E+pre-M protein heterodimers which are destroyed and reorganized by proteolytic cleavage during virus release.
    J Virol. 1989 Jun;63(6):2521-6 PMID: 2724410
  42. Antigenic relationships between flaviviruses as determined by cross-neutralization tests with polyclonal antisera.
    J Gen Virol. 1989 Jan;70 ( Pt 1):37-43 PMID: 2543738
  43. Interplay between carbohydrate in the stalk and the length of the connecting peptide determines the cleavability of influenza virus hemagglutinin.
    J Virol. 1989 Aug;63(8):3296-300 PMID: 2746732
  44. Monoclonal antibodies for dengue virus prM glycoprotein protect mice against lethal dengue infection.
    Am J Trop Med Hyg. 1989 Nov;41(5):576-80 PMID: 2817214
  45. Acidotropic amines inhibit proteolytic processing of flavivirus prM protein.
    Virology. 1990 Feb;174(2):450-8 PMID: 2154882
  46. Fusion activity of flaviviruses: comparison of mature and immature (prM-containing) tick-borne encephalitis virions.
    J Gen Virol. 1991 Jun;72 ( Pt 6):1323-9 PMID: 1710648
  47. Dengue virus premembrane and membrane proteins elicit a protective immune response.
    Virology. 1991 Nov;185(1):505-8 PMID: 1926792
  48. Comparison of a dengue-2 virus and its candidate vaccine derivative: sequence relationships with the flaviviruses and other viruses.
    Virology. 1992 Apr;187(2):573-90 PMID: 1312269
  49. Immunological analysis of human immunodeficiency virus type 1 envelope glycoprotein proteolytic cleavage.
    Virology. 1992 Apr;187(2):825-8 PMID: 1372142
  50. Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin-like endoprotease.
    EMBO J. 1992 Jul;11(7):2407-14 PMID: 1628614
  51. Purification and characterization of furin, a Kex2-like processing endoprotease, produced in Chinese hamster ovary cells.
    J Biol Chem. 1992 Aug 15;267(23):16094-9 PMID: 1644796
  52. Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen.
    J Biol Chem. 1992 Aug 15;267(23):16396-402 PMID: 1644824
  53. Anthrax toxin protective antigen is activated by a cell surface protease with the sequence specificity and catalytic properties of furin.
    Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10277-81 PMID: 1438214
  54. Inhibition of furin-mediated cleavage activation of HIV-1 glycoprotein gp160.
    Nature. 1992 Nov 26;360(6402):358-61 PMID: 1360148
  55. Expression and secretion of Japanese encephalitis virus nonstructural protein NS1 by insect cells using a recombinant baculovirus.
    Virology. 1992 Dec;191(2):826-36 PMID: 1448926
  56. The Murray Valley encephalitis virus prM protein confers acid resistance to virus particles and alters the expression of epitopes within the R2 domain of E glycoprotein.
    Virology. 1992 Dec;191(2):921-31 PMID: 1280384
  57. Processing of the dengue virus type 2 proteins prM and C-prM.
    J Gen Virol. 1993 Feb;74 ( Pt 2):175-82 PMID: 8429301
  58. Proper maturation of the Japanese encephalitis virus envelope glycoprotein requires cosynthesis with the premembrane protein.
    J Virol. 1993 Mar;67(3):1672-5 PMID: 8437237
  59. Legitimate and illegitimate cleavage of human immunodeficiency virus glycoproteins by furin.
    J Virol. 1993 Jun;67(6):3601-4 PMID: 8497067
  60. Structural changes and functional control of the tick-borne encephalitis virus glycoprotein E by the heterodimeric association with protein prM.
    Virology. 1994 Jan;198(1):109-17 PMID: 8259646
  61. The convertases furin and PC1 can both cleave the human immunodeficiency virus (HIV)-1 envelope glycoprotein gp160 into gp120 (HIV-1 SU) and gp41 (HIV-I TM).
    J Biol Chem. 1994 Apr 22;269(16):12240-7 PMID: 8163529
  62. Cleavage of proalbumin peptides by furin reveals unexpected restrictions at the P2 and P'1 sites.
    FEBS Lett. 1994 Jun 20;347(1):80-4 PMID: 8013667
  63. Activation of an influenza virus A/turkey/Oregon/71 HA insertion variant by the subtilisin-like endoprotease furin.
    Virology. 1994 Aug 1;202(2):988-91 PMID: 8030261
  64. Proprotein-processing endoproteases PC6 and furin both activate hemagglutinin of virulent avian influenza viruses.
    J Virol. 1994 Sep;68(9):6074-8 PMID: 8057485
  65. Oligomeric rearrangement of tick-borne encephalitis virus envelope proteins induced by an acidic pH.
    J Virol. 1995 Feb;69(2):695-700 PMID: 7529335
  66. Lack of augmenting effect of interferon-gamma on dengue virus multiplication in human peripheral blood monocytes.
    J Med Virol. 1995 Jan;45(1):43-9 PMID: 7536230
  67. Do hemagglutinin genes of highly pathogenic avian influenza viruses constitute unique phylogenetic lineages?
    Virology. 1995 Jun 1;209(2):664-70 PMID: 7778300
  68. Structural investigation and kinetic characterization of potential cleavage sites of HIV GP160 by human furin and PC1.
    Biochem Biophys Res Commun. 1995 Aug 4;213(1):356-61 PMID: 7639757
  69. Antibodies that block virus attachment to Vero cells are a major component of the human neutralizing antibody response against dengue virus type 2.
    J Med Virol. 1995 Apr;45(4):451-61 PMID: 7666046
  70. Characterization of Langat virus antigenic determinants defined by monoclonal antibodies to E, NS1 and preM and identification of a protective, non-neutralizing preM-specific monoclonal antibody.
    Virus Res. 1996 Aug;43(2):125-36 PMID: 8864202
  71. Structural requirements for low-pH-induced rearrangements in the envelope glycoprotein of tick-borne encephalitis virus.
    J Virol. 1996 Nov;70(11):8142-7 PMID: 8892942
  72. Identification of the paired basic convertases implicated in HIV gp160 processing based on in vitro assays and expression in CD4(+) cell lines.
    J Biol Chem. 1996 Nov 29;271(48):30442-50 PMID: 8940009
  73. The role of eukaryotic subtilisin-like endoproteases for the activation of human immunodeficiency virus glycoproteins in natural host cells.
    J Virol. 1997 Feb;71(2):1036-45 PMID: 8995623
  74. Role of hemagglutinin cleavage for the pathogenicity of influenza virus.
    Virology. 1999 May 25;258(1):1-20 PMID: 10329563
  75. Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E.
    J Virol. 1999 Jul;73(7):5605-12 PMID: 10364309
  76. Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion.
    J Virol. 1999 Jul;73(7):6104-10 PMID: 10364366
  77. The furin protease cleavage recognition sequence of Sindbis virus PE2 can mediate virion attachment to cell surface heparan sulfate.
    J Virol. 1999 Aug;73(8):6299-306 PMID: 10400721
  78. Detection of dengue virus RNA in patients after primary or secondary dengue infection by using the TaqMan automated amplification system.
    J Clin Microbiol. 1999 Aug;37(8):2543-7 PMID: 10405398
  79. Proteolytic processing in the secretory pathway.
    J Biol Chem. 1999 Jul 23;274(30):20745-8 PMID: 10409610
  80. Mutagenesis of the signal sequence of yellow fever virus prM protein: enhancement of signalase cleavage In vitro is lethal for virus production.
    J Virol. 2000 Jan;74(1):24-32 PMID: 10590087
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.78.5.2367-2381.2004
Published
2004-03-00
Pages
2367-81
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC369205
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.78.5.2367-2381.2004 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2