Genome-wide siRNA Screening at Biosafety Level 4 Reveals a Crucial Role for Fibrillarin in Henipavirus Infection

PLoS Pathogens

Paul Monaghan¹  Lin-Fa Wang¹  Celine Deffrasnes¹  John W. Lowenthal¹  Chwan Hong Foo¹  Glenn A. Marsh¹  Christina L. Rootes¹  Timothy E. Adams²  Julian Grusovin²  Michael K. Lo³  Kaylene J. Simpson⁴  S. Mark Tompkins⁴  Andrew G. D. Bean⁵  Cathryn M. Gould⁶  Cameron R. Stewart⁶ 

[1] CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia;CSIRO Manufacturing, Parkville, Victoria, Australia;Centers for Disease Control & Prevention, Viral Special Pathogens Branch, Atlanta, Georgia, United States of America;Department of Infectious Diseases, University of Georgia, Athens, Georgia, United States of America, and School of Medicine, Deakin University, Waurn Ponds, Victoria, Australia;The Sir Peter MacCallum Department of Oncology, The University of Melbourne, Melbourne, Australia;Victorian Centre for Functional Genomics, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia

关键词: Small interfering RNAs;    HeLa cells;    Henipavirus;    Hendra virus;    Respiratory infections;    Cell staining;    Transfection;    Viral replication;   

DOI  :  10.1371/journal.ppat.1005478

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY