Home LiteratureArticle Details
PMID: 10357827 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity.

The EMBO journal ·Vol. 18 ·No. 11 ·1999-06-01 ·Pages 3164-72

Ahola T, Lampio A, Auvinen P, Kääriäinen L

Abstract

The replication complexes of all positive strand RNA viruses of eukaryotes are associated with membranes. In the case of Semliki Forest virus (SFV), the main determinant of membrane attachment seems to be the virus-encoded non-structural protein NSP1, the capping enzyme of the viral mRNAs, which has guanine-7-methyltransferase and guanylyltransferase activities. We show here that both enzymatic activities of SFV NSP1 are inactivated by detergents and reactivated by anionic phospholipids, especially phosphatidylserine. The region of NSP1 responsible for binding to membranes as well as to liposomes was mapped to a short segment, which is conserved in the large alphavirus-like superfamily of viruses. A synthetic peptide of 20 amino acids from the putative binding site competed with in vitro synthesized NSP1 for binding to liposomes containing phosphatidylserine. These findings suggest a molecular mechanism by which RNA virus replicases attach to intracellular membranes and why they depend on the membranous environment.

MeSH Terms
Amino Acid Sequence Anions/metabolism Binding, Competitive Cell Membrane/chemistry,metabolism Conserved Sequence/genetics Detergents/pharmacology Enzyme Activation/drug effects Escherichia coli/cytology,genetics,metabolism Liposomes/antagonists & inhibitors,metabolism Molecular Sequence Data Mutation Nucleotidyltransferases/antagonists & inhibitors,chemistry,genetics,metabolism Peptide Fragments/chemical synthesis,metabolism Phosphatidylserines/metabolism,pharmacology Phospholipids/metabolism,pharmacology Protein Binding/drug effects Protein Biosynthesis Protein Conformation/drug effects Recombinant Proteins/analysis,biosynthesis,metabolism S-Adenosylmethionine/antagonists & inhibitors,metabolism Semliki forest virus/enzymology,genetics Solubility/drug effects Viral Nonstructural Proteins/antagonists & inhibitors,chemistry,genetics,metabolism
Chemicals
Anions Detergents Liposomes Peptide Fragments Phosphatidylserines Phospholipids Recombinant Proteins Viral Nonstructural Proteins S-Adenosylmethionine Nucleotidyltransferases mRNA guanylyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ahola T
Institute of Biotechnology, Viikki Biocenter, PO Box 56 (Viikinkaari 9), FIN-00014 University of Helsinki, Finland.
Lampio A
Auvinen P
Kääriäinen L
References (45)
45 references, click to expand
  1. Genome similarities between plant and animal RNA viruses.
    Microbiol Sci. 1987 Jul;4(7):197-202 PMID: 3153611
  2. Sindbis virus nsP1 functions in negative-strand RNA synthesis.
    J Virol. 1991 Feb;65(2):985-8 PMID: 1824787
  3. Biogenesis of type I cytopathic vacuoles in Semliki Forest virus-infected BHK cells.
    J Virol. 1991 Mar;65(3):1623-7 PMID: 1825342
  4. Regulation of CTP:phosphocholine cytidylyltransferase by lipids. 1. Negative surface charge dependence for activation.
    Biochemistry. 1991 Jun 18;30(24):5873-80 PMID: 1646002
  5. Conservation of the putative methyltransferase domain: a hallmark of the 'Sindbis-like' supergroup of positive-strand RNA viruses.
    J Gen Virol. 1992 Aug;73 ( Pt 8):2129-34 PMID: 1645151
  6. Active complete in vitro replication of nodavirus RNA requires glycerophospholipid.
    Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11136-40 PMID: 1454791
  7. Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences.
    Crit Rev Biochem Mol Biol. 1993;28(5):375-430 PMID: 8269709
  8. Properties of purified recombinant poliovirus protein 3aB as substrate for viral proteinases and as co-factor for RNA polymerase 3Dpol.
    J Biol Chem. 1994 Jan 7;269(1):66-70 PMID: 8276867
  9. The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1.
    Nature. 1994 Jan 20;367(6460):243-9 PMID: 8121489
  10. Characterization of rubella virus replication complexes using antibodies to double-stranded RNA.
    Virology. 1994 Apr;200(1):307-12 PMID: 8128633
  11. Genetic analysis of the nsP3 region of Sindbis virus: evidence for roles in minus-strand and subgenomic RNA synthesis.
    J Virol. 1994 Sep;68(9):5781-91 PMID: 8057460
  12. ATPase and GTPase activities associated with Semliki Forest virus nonstructural protein nsP2.
    J Virol. 1994 Sep;68(9):5804-10 PMID: 8057461
  13. Expression of Semliki Forest virus nsP1-specific methyltransferase in insect cells and in Escherichia coli.
    J Virol. 1994 Nov;68(11):7418-25 PMID: 7933125
  14. The alphaviruses: gene expression, replication, and evolution.
    Microbiol Rev. 1994 Sep;58(3):491-562 PMID: 7968923
  15. Reaction in alphavirus mRNA capping: formation of a covalent complex of nonstructural protein nsP1 with 7-methyl-GMP.
    Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):507-11 PMID: 7831320
  16. Membrane-binding peptide from the C2 domain of factor VIII forms an amphipathic structure as determined by NMR spectroscopy.
    Biochemistry. 1995 Mar 7;34(9):3022-31 PMID: 7893714
  17. Amino terminal regions of poliovirus 2C protein mediate membrane binding.
    Virology. 1995 Apr 20;208(2):540-53 PMID: 7747426
  18. The alphavirus replicase protein nsP1 is membrane-associated and has affinity to endocytic organelles.
    Virology. 1995 Apr 20;208(2):610-20 PMID: 7747433
  19. Mutagenesis of the Sindbis virus nsP1 protein: effects on methyltransferase activity and viral infectivity.
    Virology. 1996 Mar 15;217(2):527-31 PMID: 8610444
  20. When two strands are better than one: the mediators and modulators of the cellular responses to double-stranded RNA.
    Virology. 1996 May 15;219(2):339-49 PMID: 8638399
  21. T7 vectors with modified T7lac promoter for expression of proteins in Escherichia coli.
    Anal Biochem. 1996 May 1;236(2):371-3 PMID: 8660525
  22. Structure of the membrane binding domain of CTP:phosphocholine cytidylyltransferase.
    Biochemistry. 1996 Sep 17;35(37):11975-84 PMID: 8810902
  23. Comparison of the replication of positive-stranded RNA viruses of plants and animals.
    Adv Virus Res. 1996;47:159-251 PMID: 8895833
  24. Determinants of membrane association for poliovirus protein 3AB.
    J Biol Chem. 1996 Oct 25;271(43):26810-8 PMID: 8900162
  25. The effects of palmitoylation on membrane association of Semliki forest virus RNA capping enzyme.
    J Biol Chem. 1996 Nov 8;271(45):28567-71 PMID: 8910486
  26. Critical residues of Semliki Forest virus RNA capping enzyme involved in methyltransferase and guanylyltransferase-like activities.
    J Virol. 1997 Jan;71(1):392-7 PMID: 8985362
  27. Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism.
    J Bacteriol. 1968 Mar;95(3):833-43 PMID: 4868364
  28. Two dimensional then layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots.
    Lipids. 1970 May;5(5):494-6 PMID: 5483450
  29. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  30. Specific Sindbis virus-coded function for minus-strand RNA synthesis.
    J Virol. 1981 Aug;39(2):348-58 PMID: 7277580
  31. Solubilization of functional membrane proteins.
    Methods Enzymol. 1984;104:305-18 PMID: 6232441
  32. Complete nucleotide sequence of the nonstructural protein genes of Semliki Forest virus.
    Nucleic Acids Res. 1986 Jul 25;14(14):5667-82 PMID: 3488539
  33. Composition of octyl glucoside-phosphatidylcholine mixed micelles.
    Biochemistry. 1988 Apr 19;27(8):2839-46 PMID: 3401451
  34. Semliki Forest virus-specific non-structural protein nsP3 is a phosphoprotein.
    J Gen Virol. 1988 Sep;69 ( Pt 9):2165-78 PMID: 2970523
  35. Alphavirus RNA replicase is located on the cytoplasmic surface of endosomes and lysosomes.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2075-86 PMID: 2904446
  36. Oxidation of glycolipids in liposomes by galactose oxidase.
    Eur J Biochem. 1988 Dec 1;178(1):87-91 PMID: 3203695
  37. Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy.
    Histochem J. 1989 Mar;21(3):163-71 PMID: 2722561
  38. Site-directed mutagenesis by overlap extension using the polymerase chain reaction.
    Gene. 1989 Apr 15;77(1):51-9 PMID: 2744487
  39. Processing the nonstructural polyproteins of sindbis virus: nonstructural proteinase is in the C-terminal half of nsP2 and functions both in cis and in trans.
    J Virol. 1989 Nov;63(11):4653-64 PMID: 2529379
  40. Binding of human factor VIII to phospholipid vesicles.
    J Biol Chem. 1990 Jan 15;265(2):815-22 PMID: 2104832
  41. Nuclear localization of Semliki Forest virus-specific nonstructural protein nsP2.
    J Virol. 1990 May;64(5):1888-96 PMID: 2139138
  42. Synthetic factor VIII peptides with amino acid sequences contained within the C2 domain of factor VIII inhibit factor VIII binding to phosphatidylserine.
    Blood. 1990 May 15;75(10):1999-2004 PMID: 2110840
  43. Bio-Beads: an efficient strategy for two-dimensional crystallization of membrane proteins.
    J Struct Biol. 1997 Apr;118(3):226-35 PMID: 9169232
  44. Crystal structure of truncated human apolipoprotein A-I suggests a lipid-bound conformation.
    Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12291-6 PMID: 9356442
  45. Poliovirus 2C protein determinants of membrane binding and rearrangements in mammalian cells.
    J Virol. 1997 Dec;71(12):8962-72 PMID: 9371552
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/18.11.3164
Published
1999-06-01
Pages
3164-72
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171397
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/18.11.3164 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: [email protected]

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