Home LiteratureArticle Details
PMID: 7628436 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A human virus protein, poliovirus protein 2BC, induces membrane proliferation and blocks the exocytic pathway in the yeast Saccharomyces cerevisiae.

The EMBO journal ·Vol. 14 ·No. 14 ·1995-07-17 ·Pages 3349-64

Barco A, Carrasco L

Abstract

Inducible synthesis of poliovirus protein 2BC in Saccharomyces cerevisiae arrests cell growth in the G2 phase of the cell cycle, while no effects are observed upon expression of poliovirus genes 2B or 2C, either individually or in combination. Expression of 2BC induces a number of morphological modifications in yeast cells, one of the most striking being the proliferation of small membranous vesicles that fill most of the cytoplasm. These vesicles are morphologically similar to the cytopathic vacuoles that proliferate during the infection of human cells by poliovirus. The transport and processing of several yeast proteins, including vacuolar carboxypeptidase Y, aminopeptidase I or yeast alpha-mating factor, is hampered upon expression of poliovirus 2BC, suggesting that transport of proteins through the Golgi apparatus is impaired by this viral protein. Finally, a number of 2BC variants were generated and the effects of their expression on yeast growth, cellular morphology and protein processing were analyzed. 2BC variants defective in the NTPase activity were still able to interfere with yeast growth and the exocytic system, while deletion of 30 amino acids at the N-terminus of 2BC impairs its function. These findings lend support to the idea that 2BC, but not 2B or 2C, is the protein responsible for vesicle proliferation in poliovirus-infected cells. In addition, the activity of a human virus protein in yeast cells opens new avenues to investigate the exact location at which poliovirus 2BC interferes with the vesicular system and to test the action of other animal virus proteins potentially involved in modifying the vesicular system in mammalian cells.

MeSH Terms
Base Sequence Biological Transport Carrier Proteins/genetics,physiology Cloning, Molecular Exocytosis/physiology HeLa Cells Humans Intracellular Membranes/metabolism Molecular Sequence Data Poliovirus/genetics,physiology Protein Processing, Post-Translational Recombinant Proteins/genetics,metabolism Saccharomyces cerevisiae/genetics,growth & development,metabolism Viral Nonstructural Proteins/genetics,physiology Virus Replication/physiology
Chemicals
2BC protein, Poliovirus Carrier Proteins Recombinant Proteins Viral Nonstructural Proteins 2C protein, viral
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Barco A
Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma de Madrid, Spain.
Carrasco L
References (50)
50 references, click to expand
  1. Membranous structures associated with translation and transcription of poliovirus RNA.
    Science. 1969 Nov 14;166(3907):885-6 PMID: 4310189
  2. Genetics of poliovirus.
    Annu Rev Genet. 1993;27:353-436 PMID: 8122908
  3. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway.
    Cell. 1980 Aug;21(1):205-15 PMID: 6996832
  4. Intracellular distribution of poliovirus proteins and the induction of virus-specific cytoplasmic structures.
    Virology. 1983 Nov;131(1):39-48 PMID: 6316654
  5. Glycosylation and processing of prepro-alpha-factor through the yeast secretory pathway.
    Cell. 1984 Feb;36(2):309-18 PMID: 6420074
  6. Two nuclear mutations that block mitochondrial protein import in yeast.
    Proc Natl Acad Sci U S A. 1984 Aug;81(15):4819-23 PMID: 6235522
  7. Association of poliovirus proteins with the endoplasmic reticulum.
    J Virol. 1984 Dec;52(3):777-83 PMID: 6092710
  8. Genetic complementation among poliovirus mutants derived from an infectious cDNA clone.
    J Virol. 1986 Dec;60(3):1040-9 PMID: 3023656
  9. GTP-binding domain: three consensus sequence elements with distinct spacing.
    Proc Natl Acad Sci U S A. 1987 Apr;84(7):1814-8 PMID: 3104905
  10. The ras-like yeast YPT1 gene is itself essential for growth, sporulation, and starvation response.
    Mol Cell Biol. 1987 Jul;7(7):2367-77 PMID: 3302675
  11. Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi.
    Annu Rev Biochem. 1987;56:829-52 PMID: 3304148
  12. Antagonistic controls regulate copy number of the yeast 2 mu plasmid.
    EMBO J. 1987 Dec 20;6(13):4205-12 PMID: 2832156
  13. Cloning of the orotidine 5'-phosphate decarboxylase (ODC) gene of Schwanniomyces occidentalis by complementation of the ura3 mutation in S. cerevisiae.
    Curr Genet. 1988;13(1):29-35 PMID: 2834102
  14. Increased amounts of HMG-CoA reductase induce "karmellae": a proliferation of stacked membrane pairs surrounding the yeast nucleus.
    J Cell Biol. 1988 Jul;107(1):101-14 PMID: 3292536
  15. Organelle assembly in yeast: characterization of yeast mutants defective in vacuolar biogenesis and protein sorting.
    J Cell Biol. 1988 Oct;107(4):1369-83 PMID: 3049619
  16. The dynamic nature of the Golgi complex.
    J Cell Biol. 1989 Feb;108(2):277-97 PMID: 2537312
  17. The Saccharomyces cerevisiae SEC14 gene encodes a cytosolic factor that is required for transport of secretory proteins from the yeast Golgi complex.
    J Cell Biol. 1989 Apr;108(4):1271-81 PMID: 2466847
  18. KAR2, a karyogamy gene, is the yeast homolog of the mammalian BiP/GRP78 gene.
    Cell. 1989 Jun 30;57(7):1211-21 PMID: 2661018
  19. An intragenic revertant of a poliovirus 2C mutant has an uncoating defect.
    J Virol. 1990 Mar;64(3):1102-7 PMID: 2154595
  20. Distinct sets of SEC genes govern transport vesicle formation and fusion early in the secretory pathway.
    Cell. 1990 May 18;61(4):723-33 PMID: 2188733
  21. Phospholipid biosynthesis and poliovirus genome replication, two coupled phenomena.
    EMBO J. 1990 Jun;9(6):2011-6 PMID: 2161336
  22. Poliovirus genetics.
    Curr Top Microbiol Immunol. 1990;161:155-88 PMID: 2169383
  23. Product review. New mammalian expression vectors.
    Nature. 1990 Nov 1;348(6296):91-2 PMID: 2234068
  24. A novel pathway of import of alpha-mannosidase, a marker enzyme of vacuolar membrane, in Saccharomyces cerevisiae.
    J Biol Chem. 1990 Dec 25;265(36):22418-25 PMID: 2266133
  25. Methods for studying the yeast vacuole.
    Methods Enzymol. 1991;194:644-61 PMID: 1706462
  26. Proteins involved in vesicular transport and membrane fusion.
    Curr Opin Cell Biol. 1991 Aug;3(4):615-20 PMID: 1772655
  27. Molecular dissection of the secretory pathway.
    Nature. 1992 Jan 30;355(6359):409-15 PMID: 1734280
  28. Human p53 and CDC2Hs genes combine to inhibit the proliferation of Saccharomyces cerevisiae.
    Mol Cell Biol. 1992 Mar;12(3):1357-65 PMID: 1545817
  29. Inhibition of poliovirus RNA synthesis by brefeldin A.
    J Virol. 1992 Apr;66(4):1985-94 PMID: 1312615
  30. Structural and functional characterization of the poliovirus replication complex.
    J Virol. 1992 May;66(5):2740-7 PMID: 1313898
  31. Vesicle-mediated protein sorting.
    Annu Rev Biochem. 1992;61:471-516 PMID: 1497318
  32. Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway.
    J Cell Biol. 1992 Oct;119(2):287-99 PMID: 1400574
  33. Intracellular localization of poliovirus RNA by in situ hybridization at the ultrastructural level using single-stranded riboprobes.
    Virology. 1992 Dec;191(2):687-97 PMID: 1333118
  34. The VPS16 gene product associates with a sedimentable protein complex and is essential for vacuolar protein sorting in yeast.
    J Biol Chem. 1993 Mar 5;268(7):4953-62 PMID: 8444873
  35. Brefeldin A reversibly blocks early but not late protein transport steps in the yeast secretory pathway.
    EMBO J. 1993 Mar;12(3):869-77 PMID: 8458343
  36. Poliovirus protein 2C has ATPase and GTPase activities.
    J Biol Chem. 1993 Apr 15;268(11):8105-10 PMID: 8385138
  37. Assembly of vaccinia virus: role of the intermediate compartment between the endoplasmic reticulum and the Golgi stacks.
    J Cell Biol. 1993 May;121(3):521-41 PMID: 8486734
  38. Membrane rearrangement and vesicle induction by recombinant poliovirus 2C and 2BC in human cells.
    Virology. 1994 Jul;202(1):129-45 PMID: 8009827
  39. Hybrid proteins between Pseudomonas exotoxin A and poliovirus protease 2Apro.
    FEBS Lett. 1994 Nov 21;355(1):45-8 PMID: 7957960
  40. Physical interaction of the HIV-1 Nef protein with beta-COP, a component of non-clathrin-coated vesicles essential for membrane traffic.
    J Biol Chem. 1994 Dec 2;269(48):30073-6 PMID: 7982906
  41. Induction of membrane proliferation by poliovirus proteins 2C and 2BC.
    Biochem Biophys Res Commun. 1995 Jan 5;206(1):64-76 PMID: 7818552
  42. Poliovirus protein 2C contains two regions involved in RNA binding activity.
    J Biol Chem. 1995 Apr 28;270(17):10105-12 PMID: 7730315
  43. Cloning and inducible synthesis of poliovirus non-structural proteins in Saccharomyces cerevisiae.
    Gene. 1995 Apr 14;156(1):19-25 PMID: 7737511
  44. Modification of membrane permeability by animal viruses.
    Adv Virus Res. 1995;45:61-112 PMID: 7793329
  45. ELECTRON MICROSCOPIC STUDY OF THE FORMATION OF POLIOVIRUS.
    Virology. 1965 Jul;26:379-89 PMID: 14319710
  46. A poliovirus minireplicon containing an inactive 2A proteinase is expressed in vaccinia virus-infected cells.
    J Virol. 1993 Aug;67(8):4621-9 PMID: 8392603
  47. Expression of p60v-src in Saccharomyces cerevisiae results in elevation of p34CDC28 kinase activity and release of the dependence of DNA replication on mitosis.
    Mol Cell Biol. 1993 Aug;13(8):5112-21 PMID: 7687746
  48. Brefeldin A blocks protein glycosylation and RNA replication of vesicular stomatitis virus.
    FEBS Lett. 1993 Dec 28;336(3):496-500 PMID: 8282118
  49. Studies of a putative amphipathic helix in the N-terminus of poliovirus protein 2C.
    Virology. 1994 Feb 15;199(1):188-99 PMID: 8116242
  50. Association of the polioviral RNA polymerase complex with phospholipid membranes.
    J Virol. 1976 Aug;19(2):457-66 PMID: 183011
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-07-17
Pages
3349-64
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394402
Subset
IM
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