title: Alphavirus genome delivery occurs directly at the plasma membrane in a time- and temperature-dependent process.
abstract: It is widely held that arboviruses such as the alphavirus Sindbis virus gain entry into cells by a process of receptor-mediated endocytosis followed by membrane fusion in the acid environment of the endosome. We have used an approach of direct observation of Sindbis virus entry into cells by electron microscopy and immunolabeling of virus proteins with antibodies conjugated to gold beads. We found that upon attaching to the cell surface, intact RNA-containing viruses became empty shells that could be identified only by antibody labeling. We found that the rate at which full particles were converted to empty particles increased with time and temperature. We found that this entry event takes place at temperatures that inhibit both endosome formation and membrane fusion. We conclude that entry of alphaviruses is by direct penetration of cell plasma membranes through a pore structure formed by virus and, possibly, host proteins.
authors: Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, North Carolina, USA.
authors: Ricardo
authors: R
authors: Vancini
authors: Gongbo
authors: G
authors: Wang
authors: Davis
authors: D
authors: Ferreira
authors: Raquel
authors: R
authors: Hernandez
authors: Dennis T
authors: DT
authors: Brown
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000821:virology*; D002478:Cells, Cultured; D006224:Cricetinae; D016253:Microscopy, Immunoelectron; D012367:RNA, Viral / Q000378:metabolism*; D012845:Sindbis Virus / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature; D013997:Time Factors; D053586:Virus Internalization
chemical_list: D012367:RNA, Viral
references: J Virol. 2003 Dec;77(23):12710-9
references: 14610193
references: Nat Rev Microbiol. 2004 Feb;2(2):109-22
references: 15043007
references: J Virol. 1999 Oct;73(10):8476-84
references: 10482600
references: J Virol. 1981 Mar;37(3):1060-5
references: 6262524
references: Annu Rev Cell Dev Biol. 2000;16:557-89
references: 11031247
references: J Virol. 1984 Dec;52(3):857-64
references: 6492263
references: Adv Virus Res. 1995;45:113-51
references: 7793323
references: PLoS Biol. 2007 Jul;5(7):e183
references: 17622193
references: Virol J. 2011 Jun 15;8:304
references: 21676248
references: PLoS One. 2010 Jul 08;5(7):e11479
references: 20628602
references: J Virol. 2009 Sep;83(18):9113-21
references: 19587056
references: Curr Protoc Microbiol. 2010 Feb;Chapter 15:Unit15B.1
references: 20131223
references: J Cell Biol. 1995 Nov;131(3):679-91
references: 7593189
references: Ciba Found Symp. 1982;(92):59-76
references: 6129957
references: EMBO J. 1998 Aug 17;17(16):4585-93
references: 9707418
references: J Gen Virol. 2003 Jul;84(Pt 7):1711-1721
references: 12810864
references: J Gen Virol. 1982 Jan;58 Pt 1:47-61
references: 7142969
references: Virology. 2005 Feb 5;332(1):307-15
references: 15661162
references: J Virol. 1976 Sep;19(3):775-81
references: 987251
references: FEBS Lett. 2002 Jun 19;521(1-3):62-6
references: 12067727
references: Adv Exp Med Biol. 2012;726:181-99
references: 22297514
references: J Virol. 1999 May;73(5):4272-8
references: 10196324
references: Virology. 1982 Apr 30;118(2):401-10
references: 7090184
references: Virology. 2013 Jan 20;435(2):406-14
references: 23099205
references: J Pharmacol Toxicol Methods. 2010 Mar-Apr;61(2):127-35
references: 20035892
references: Virology. 2007 Jun 5;362(2):461-7
references: 17289103
references: J Gen Virol. 2003 Jan;84(Pt 1):173-181
references: 12533714
references: J Virol. 2001 Feb;75(4):2010-3
references: 11160702
references: Mol Biol Med. 1990 Feb;7(1):17-31
references: 2182968
references: J Gen Virol. 1986 Feb;67 ( Pt 2):377-80
references: 3944588
references: J Virol. 2011 Feb;85(3):1257-66
references: 21084471
references: Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9095-9
references: 8415660
references: J Virol. 2000 Feb;74(3):1342-54
references: 10627545
references: Curr Protoc Microbiol. 2005 Oct;Chapter 15:Unit 15B.1
references: 18770557
references: Virology. 2004 Jul 1;324(2):373-86
references: 15207623
references: J Virol. 2010 Nov;84(21):10946-55
references: 20668081
references: J Virol. 1996 May;70(5):2757-63
references: 8627749
references: Virology. 2011 Jul 5;415(2):132-40
references: 21561635