title: Genome-wide siRNA Screening at Biosafety Level 4 Reveals a Crucial Role for Fibrillarin in Henipavirus Infection.
abstract: Hendra and Nipah viruses (genus Henipavirus, family Paramyxoviridae) are highly pathogenic bat-borne viruses. The need for high biocontainment when studying henipaviruses has hindered the development of therapeutics and knowledge of the viral infection cycle. We have performed a genome-wide siRNA screen at biosafety level 4 that identified 585 human proteins required for henipavirus infection. The host protein with the largest impact was fibrillarin, a nucleolar methyltransferase that was also required by measles, mumps and respiratory syncytial viruses for infection. While not required for cell entry, henipavirus RNA and protein syntheses were greatly impaired in cells lacking fibrillarin, indicating a crucial role in the RNA replication phase of infection. During infection, the Hendra virus matrix protein co-localized with fibrillarin in cell nucleoli, and co-associated as a complex in pulldown studies, while its nuclear import was unaffected in fibrillarin-depleted cells. Mutagenesis studies showed that the methyltransferase activity of fibrillarin was required for henipavirus infection, suggesting that this enzyme could be targeted therapeutically to combat henipavirus infections.
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Celine
authors.initials: C
authors.lastname: Deffrasnes
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Glenn A
authors.initials: GA
authors.lastname: Marsh
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Chwan Hong
authors.initials: CH
authors.lastname: Foo
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Christina L
authors.initials: CL
authors.lastname: Rootes
authors.affiliation: Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
authors.forename: Cathryn M
authors.initials: CM
authors.lastname: Gould
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: Julian
authors.initials: J
authors.lastname: Grusovin
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Paul
authors.initials: P
authors.lastname: Monaghan
authors.affiliation: Centers for Disease Control & Prevention, Viral Special Pathogens Branch, Atlanta, Georgia, United States of America.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Lo
authors.affiliation: Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America, and School of Medicine, Deakin University, Waurn Ponds, Victoria, Australia.
authors.forename: S Mark
authors.initials: SM
authors.lastname: Tompkins
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Adams
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.|Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America, and School of Medicine, Deakin University, Waurn Ponds, Victoria, Australia.
authors.forename: John W
authors.initials: JW
authors.lastname: Lowenthal
authors.affiliation: Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.|The Sir Peter MacCallum Department of Oncology, The University of Melbourne, Melbourne, Australia.
authors.forename: Kaylene J
authors.initials: KJ
authors.lastname: Simpson
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Cameron R
authors.initials: CR
authors.lastname: Stewart
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Andrew G D
authors.initials: AG
authors.lastname: Bean
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.|Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
authors.forename: Lin-Fa
authors.initials: LF
authors.lastname: Wang
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D045404:Hendra Virus / Q000378:metabolism; D045464:Henipavirus Infections / Q000821:virology*; D006801:Humans; D009154:Mutation; D045405:Nipah Virus / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D034741:RNA, Small Interfering; D014709:Vero Cells; D014763:Viral Matrix Proteins / Q000378:metabolism
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D034741:RNA, Small Interfering; D014763:Viral Matrix Proteins; C047458:fibrillarin