title: Recruitment of cellular clathrin to viral factories and disruption of clathrin-dependent trafficking.
abstract: The viral factories of mammalian reovirus (MRV) are cytoplasmic structures that serve as sites of viral genome replication and particle assembly. A 721-aa MRV non-structural protein, µNS, forms the factory matrix and recruits other viral proteins to these structures. In this report, we show that µNS contains a conserved C-proximal sequence (711-LIDFS-715) that is similar to known clathrin-box motifs and is required for recruitment of clathrin to viral factories. Clathrin recruitment by µNS occurs independently of infecting MRV particles or other MRV proteins. Ala substitution for a single Leu residue (mutation L711A) within the putative clathrin-binding motif of µNS inhibits clathrin recruitment, but does not prevent formation or expansion of viral factories. Notably, clathrin-dependent cellular functions, including both endocytosis and secretion, are disrupted in cells infected with MRV expressing wild-type, but not L711A, µNS. These results identify µNS as a novel adaptor-like protein that recruits cellular clathrin to viral factories, disrupting normal functions of clathrin in cellular membrane trafficking. To our knowledge, this is the only viral or bacterial protein yet shown to interfere with clathrin functions in this manner. The results additionally establish a new approach for studies of clathrin functions, based on µNS-mediated sequestration.
authors: Department of Microbiology & Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors: Tijana
authors: T
authors: Ivanovic
authors: Steeve
authors: S
authors: Boulant
authors: Marcelo
authors: M
authors: Ehrlich
authors: Aleksander A
authors: AA
authors: Demidenko
authors: Michelle M
authors: MM
authors: Arnold
authors: Tomas
authors: T
authors: Kirchhausen
authors: Max L
authors: ML
authors: Nibert
mesh_terms: D033961:Adaptor Protein Complex 1 / Q000235:genetics / Q000378:metabolism; D033962:Adaptor Protein Complex 2 / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002966:Clathrin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism; D007181:Inclusion Bodies, Viral / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D030102:Orthoreovirus, Mammalian / Q000472:pathogenicity / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012088:Reoviridae Infections / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D033961:Adaptor Protein Complex 1; D033962:Adaptor Protein Complex 2; D002966:Clathrin; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins