title: Rotavirus increases levels of lipidated LC3 supporting accumulation of infectious progeny virus without inducing autophagosome formation.
abstract: Replication of many RNA viruses benefits from subversion of the autophagic pathway through many different mechanisms. Rotavirus, the main etiologic agent of pediatric gastroenteritis worldwide, has been recently described to induce accumulation of autophagosomes as a mean for targeting viral proteins to the sites of viral replication. Here we show that the viral-induced increase of the lipidated form of LC3 does not correlate with an augmented formation of autophagosomes, as detected by immunofluorescence and electron microscopy. The LC3-II accumulation was found to be dependent on active rotavirus replication through the use of antigenically intact inactivated viral particles and of siRNAs targeting viral genes that are essential for viral replication. Silencing expression of LC3 or of Atg7, a protein involved in LC3 lipidation, resulted in a significant impairment of viral titers, indicating that these elements of the autophagic pathway are required at late stages of the viral cycle.
authors.affiliation: Department of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, Italy; International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano (Trieste), Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Arnoldi
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano (Trieste), Italy.
authors.forename: Giuditta
authors.initials: G
authors.lastname: De Lorenzo
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano (Trieste), Italy.
authors.forename: Miguel
authors.initials: M
authors.lastname: Mano
authors.affiliation: Institute of Veterinary Anatomy, University of Zürich, Zürich, Switzerland; Institute of Virology, University of Zürich, Zürich, Switzerland.
authors.forename: Elisabeth M
authors.initials: EM
authors.lastname: Schraner
authors.affiliation: Institute of Veterinary Anatomy, University of Zürich, Zürich, Switzerland; Institute of Virology, University of Zürich, Zürich, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Wild
authors.affiliation: Institute of Virology, University of Zürich, Zürich, Switzerland.
authors.forename: Catherine
authors.initials: C
authors.lastname: Eichwald
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano (Trieste), Italy.
authors.forename: Oscar R
authors.initials: OR
authors.lastname: Burrone
mesh_terms: D000818:Animals; D001343:Autophagy; D002460:Cell Line; D002522:Chlorocebus aethiops; D050356:Lipid Metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism*; D012401:Rotavirus / Q000502:physiology*; D014779:Virus Replication
chemical_list: D008869:Microtubule-Associated Proteins
references.citation: Virology. 2011 Feb 5;410(1):142-50
references.pmid: 21112602
references.citation: Methods Mol Med. 2000;34:33-66
references.pmid: 21318854
references.citation: J Gen Virol. 1987 Jan;68 ( Pt 1):115-22
references.pmid: 3027239
references.citation: EMBO Rep. 2008 Sep;9(9):859-64
references.pmid: 18704115
references.citation: Autophagy. 2007 Jul-Aug;3(4):323-8
references.pmid: 17387262
references.citation: Nature. 2013 Feb 14;494(7436):201-6
references.pmid: 23364696
references.citation: Proc Natl Acad Sci U S A. 2011 Mar 22;108(12):4834-9
references.pmid: 21383145
references.citation: J Gen Virol. 1991 Jul;72 ( Pt 7):1551-7
references.pmid: 1713260
references.citation: Autophagy. 2007 Nov-Dec;3(6):586-90
references.pmid: 17786021
references.citation: Semin Immunopathol. 2010 Dec;32(4):323-41
references.pmid: 20865416
references.citation: Curr Opin Virol. 2012 Aug;2(4):443-8
references.pmid: 22698800
references.citation: J Cell Biol. 2000 Oct 16;151(2):263-76
references.pmid: 11038174
references.citation: Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1997 Feb;19(1):48-53
references.pmid: 10453552
references.citation: J Virol. 1979 Sep;31(3):810-5
references.pmid: 229253
references.citation: J Gen Virol. 2004 Mar;85(Pt 3):625-634
references.pmid: 14993647
references.citation: Science. 2010 Dec 3;330(6009):1344-8
references.pmid: 21127245
references.citation: Virology. 1991 May;182(1):8-16
references.pmid: 1850914
references.citation: J Gen Virol. 2004 Nov;85(Pt 11):3285-3290
references.pmid: 15483242
references.citation: PLoS One. 2012;7(10):e47947
references.pmid: 23110139
references.citation: Autophagy. 2013 Feb 1;9(2):164-74
references.pmid: 23182945
references.citation: J Gen Virol. 2005 May;86(Pt 5):1481-1487
references.pmid: 15831961
references.citation: J Virol. 1989 Nov;63(11):4553-62
references.pmid: 2552139
references.citation: J Virol. 2002 Apr;76(7):3461-70
references.pmid: 11884570
references.citation: Autophagy. 2010 Feb;6(2):217-27
references.pmid: 20104022
references.citation: Proc Natl Acad Sci U S A. 2012 Dec 11;109(50):E3405-13
references.pmid: 23184977
references.citation: J Virol. 2006 Jun;80(12):6061-71
references.pmid: 16731945
references.citation: J Virol. 2011 Mar;85(6):2781-92
references.pmid: 21228236
references.citation: Cell Host Microbe. 2010 Jun 25;7(6):500-8
references.pmid: 20542253
references.citation: J Virol. 2010 Dec;84(23):12110-24
references.pmid: 20861268
references.citation: J Gen Virol. 1985 Dec;66 ( Pt 12):2715-24
references.pmid: 2999314
references.citation: J Virol. 1997 Nov;71(11):8268-78
references.pmid: 9343179
references.citation: J Cell Biol. 2009 Dec 14;187(6):875-88
references.pmid: 20008565
references.citation: Nat Rev Mol Cell Biol. 2007 Sep;8(9):741-52
references.pmid: 17717517
references.citation: Virology. 2010 Sep 15;405(1):1-7
references.pmid: 20580051
references.citation: J Gen Virol. 1998 Nov;79 ( Pt 11):2679-86
references.pmid: 9820143
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):14046-51
references.pmid: 19666601
references.citation: J Virol Methods. 1992 Jul;38(1):93-102
references.pmid: 1322935
references.citation: J Gen Virol. 1996 Sep;77 ( Pt 9):2059-65
references.pmid: 8811003
references.citation: Int J Biochem Cell Biol. 2012 Jun;44(6):989-97
references.pmid: 22465708
references.citation: Microbes Infect. 2012 Feb;14(2):126-39
references.pmid: 22051604
references.citation: J Gen Virol. 2005 Jun;86(Pt 6):1609-1617
references.pmid: 15914838
references.citation: FEBS Lett. 2010 Apr 2;584(7):1287-95
references.pmid: 20083114
references.citation: J Med Virol. 2009 Jul;81(7):1241-52
references.pmid: 19475621
references.citation: J Virol. 2008 Sep;82(18):9143-53
references.pmid: 18596087
references.citation: J Virol. 2007 Mar;81(5):2128-37
references.pmid: 17182692
references.citation: Annu Rev Biochem. 2011;80:125-56
references.pmid: 21548784
references.citation: EMBO J. 2000 Nov 1;19(21):5720-8
references.pmid: 11060023
references.citation: Lancet Infect Dis. 2012 Feb;12(2):136-41
references.pmid: 22030330
references.citation: Nat Cell Biol. 2009 Apr;11(4):385-96
references.pmid: 19270696
references.citation: Autophagy. 2005 Jul;1(2):84-91
references.pmid: 16874052