title: Complex dynamic development of poliovirus membranous replication complexes.
abstract: Replication of all positive-strand RNA viruses is intimately associated with membranes. Here we utilize electron tomography and other methods to investigate the remodeling of membranes in poliovirus-infected cells. We found that the viral replication structures previously described as "vesicles" are in fact convoluted, branching chambers with complex and dynamic morphology. They are likely to originate from cis-Golgi membranes and are represented during the early stages of infection by single-walled connecting and branching tubular compartments. These early viral organelles gradually transform into double-membrane structures by extension of membranous walls and/or collapsing of the luminal cavity of the single-membrane structures. As the double-membrane regions develop, they enclose cytoplasmic material. At this stage, a continuous membranous structure may have double- and single-walled membrane morphology at adjacent cross-sections. In the late stages of the replication cycle, the structures are represented mostly by double-membrane vesicles. Viral replication proteins, double-stranded RNA species, and actively replicating RNA are associated with both double- and single-membrane structures. However, the exponential phase of viral RNA synthesis occurs when single-membrane formations are predominant in the cell. It has been shown previously that replication complexes of some other positive-strand RNA viruses form on membrane invaginations, which result from negative membrane curvature. Our data show that the remodeling of cellular membranes in poliovirus-infected cells produces structures with positive curvature of membranes. Thus, it is likely that there is a fundamental divergence in the requirements for the supporting cellular membrane-shaping machinery among different groups of positive-strand RNA viruses.
authors.affiliation: Virginia-Maryland College of Veterinary Medicine, University of Maryland, College Park, Maryland 20742, USA. gbelov@umd.edu
authors.forename: George A
authors.initials: GA
authors.lastname: Belov
authors.forename: Vinod
authors.initials: V
authors.lastname: Nair
authors.forename: Bryan T
authors.initials: BT
authors.lastname: Hansen
authors.forename: Forrest H
authors.initials: FH
authors.lastname: Hoyt
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Fischer
authors.forename: Ellie
authors.initials: E
authors.lastname: Ehrenfeld
mesh_terms: D002462:Cell Membrane / Q000378:metabolism / Q000821:virology; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000821:virology; D006056:Golgi Apparatus / Q000378:metabolism / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism / Q000821:virology*; D011051:Poliomyelitis / Q000378:metabolism / Q000821:virology*; D017955:Poliovirus / Q000235:genetics / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication
chemical_list: D014764:Viral Proteins
references.citation: PLoS Biol. 2005 May;3(5):e156
references.pmid: 15884975
references.citation: Cell Microbiol. 2010 Sep 1;12(9):1235-49
references.pmid: 20331642
references.citation: J Virol. 2009 Jul;83(13):6599-609
references.pmid: 19369338
references.citation: J Virol. 2007 Sep;81(17):9259-67
references.pmid: 17567696
references.citation: PLoS Biol. 2008 Sep 16;6(9):e226
references.pmid: 18798692
references.citation: J Virol. 2007 Jan;81(2):558-67
references.pmid: 17079330
references.citation: J Virol. 1992 Apr;66(4):1985-94
references.pmid: 1312615
references.citation: J Virol. 1996 Dec;70(12):8675-83
references.pmid: 8970994
references.citation: J Virol. 2003 Dec;77(23):12819-28
references.pmid: 14610203
references.citation: J Biol Chem. 1994 Jan 7;269(1):66-70
references.pmid: 8276867
references.citation: Microbiol Sci. 1987 Jul;4(7):197-202
references.pmid: 3153611
references.citation: Cell. 2010 May 14;141(4):656-67
references.pmid: 20478256
references.citation: Nat Cell Biol. 2009 Dec;11(12):1433-7
references.pmid: 19898463
references.citation: J Virol. 1996 Oct;70(10):6576-88
references.pmid: 8794292
references.citation: J Virol. 1972 Sep;10(3):492-503
references.pmid: 4672392
references.citation: J Virol. 1985 Feb;53(2):536-42
references.pmid: 3968720
references.citation: Autophagy. 2009 Nov;5(8):1180-5
references.pmid: 19855179
references.citation: Curr Top Microbiol Immunol. 2005;285:139-73
references.pmid: 15609503
references.citation: Cell. 2010 May 28;141(5):799-811
references.pmid: 20510927
references.citation: Crit Rev Biochem Mol Biol. 1993;28(5):375-430
references.pmid: 8269709
references.citation: Autophagy. 2008 Apr;4(3):286-9
references.pmid: 18094610
references.citation: Cell Host Microbe. 2009 Apr 23;5(4):365-75
references.pmid: 19380115
references.citation: J Virol. 2011 May;85(9):4284-96
references.pmid: 21345960
references.citation: Nature. 1993 Aug 26;364(6440):771-6
references.pmid: 8102790
references.citation: J Cell Biol. 1999 Nov 29;147(5):945-58
references.pmid: 10579716
references.citation: Biochim Biophys Acta. 1971 Jun 1;233(3):504-12
references.pmid: 5165434
references.citation: PLoS Pathog. 2008 Nov;4(11):e1000216
references.pmid: 19023417
references.citation: Virology. 1964 Jun;23:252-8
references.pmid: 14187912
references.citation: Virology. 1993 Jun;194(2):878-81
references.pmid: 8503193
references.citation: Nat Rev Microbiol. 2008 Dec;6(12):925-39
references.pmid: 18997823
references.citation: Trends Cell Biol. 1999 Feb;9(2):81-5
references.pmid: 10087625
references.citation: Annu Rev Biochem. 2005;74:833-65
references.pmid: 15952904
references.citation: J Virol. 2007 Oct;81(19):10815-7
references.pmid: 17670838
references.citation: J Virol. 2005 Apr;79(8):5163-73
references.pmid: 15795300
references.citation: J Biol Chem. 2002 Oct 25;277(43):40434-41
references.pmid: 12183456
references.citation: J Cell Biol. 1962 Aug;14:281-302
references.pmid: 13883195
references.citation: J Biophys Biochem Cytol. 1958 May 25;4(3):301-8
references.pmid: 13549502
references.citation: J Gen Virol. 2005 Mar;86(Pt 3):707-718
references.pmid: 15722531
references.citation: J Virol. 2010 Aug;84(15):7543-57
references.pmid: 20484502
references.citation: Virology. 1972 Jan;47(1):39-47
references.pmid: 4333576
references.citation: Virology. 1980 Jan 30;100(2):390-9
references.pmid: 6243433
references.citation: Virology. 1980 Mar;101(2):389-98
references.pmid: 6244696
references.citation: Virology. 1968 Oct;36(2):241-53
references.pmid: 4879189
references.citation: J Cell Biol. 1995 Dec;131(6 Pt 2):1715-26
references.pmid: 8557739
references.citation: J Virol. 1992 May;66(5):2740-7
references.pmid: 1313898
references.citation: EMBO J. 1997 Jun 16;16(12):3519-32
references.pmid: 9218794
references.citation: J Virol. 2000 Oct;74(19):8953-65
references.pmid: 10982339
references.citation: Methods Mol Biol. 2009;510:195-209
references.pmid: 19009263
references.citation: Virology. 1983 Nov;131(1):39-48
references.pmid: 6316654
references.citation: J Virol. 2008 Sep;82(18):9143-53
references.pmid: 18596087
references.citation: Virology. 1967 Jun;32(2):201-15
references.pmid: 4290640
references.citation: Cell Microbiol. 2010 Oct;12(10):1463-79
references.pmid: 20497182
references.citation: Virology. 1992 Nov;191(1):166-75
references.pmid: 1329315
references.citation: PLoS Biol. 2007 Sep;5(9):e220
references.pmid: 17696647
references.citation: J Virol. 2007 Nov;81(22):12543-53
references.pmid: 17804493
references.citation: Virology. 1987 Sep;160(1):220-6
references.pmid: 2820130
references.citation: J Virol. 2001 Oct;75(20):9808-18
references.pmid: 11559814
references.citation: Biken J. 1977 Jun;20(2):57-67
references.pmid: 412491
references.citation: J Virol. 2002 Oct;76(19):9856-67
references.pmid: 12208963
references.citation: J Microsc. 1974 Sep;102(Pt 1):29-39
references.pmid: 4216636