title: Assembly of subtype 1 influenza neuraminidase is driven by both the transmembrane and head domains.
abstract: Neuraminidase (NA) is one of the two major influenza surface antigens and the main influenza drug target. Although NA has been well characterized and thought to function as a tetramer, the role of the transmembrane domain (TMD) in promoting proper NA assembly has not been systematically studied. Here, we demonstrate that in the absence of the TMD, NA is synthesized and transported in a predominantly inactive state. Substantial activity was rescued by progressive truncations of the stalk domain, suggesting the TMD contributes to NA maturation by tethering the stalk to the membrane. To analyze how the TMD supports NA assembly, the TMD was examined by itself. The NA TMD formed a homotetramer and efficiently trafficked to the plasma membrane, indicating the TMD and enzymatic head domain drive assembly together through matching oligomeric states. In support of this, an unrelated strong oligomeric TMD rescued almost full NA activity, whereas the weak oligomeric mutant of this TMD restored only half of wild type activity. These data illustrate that a large soluble domain can force assembly with a poorly compatible TMD; however, optimal assembly requires coordinated oligomerization between the TMD and the soluble domain.
authors.affiliation: Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Diogo V
authors.initials: DV
authors.lastname: da Silva
authors.forename: Johan
authors.initials: J
authors.lastname: Nordholm
authors.forename: Ursula
authors.initials: U
authors.lastname: Madjo
authors.forename: Annika
authors.initials: A
authors.lastname: Pfeiffer
authors.forename: Robert
authors.initials: R
authors.lastname: Daniels
mesh_terms: D000818:Animals; D019281:Dimerization; D004285:Dogs; D006023:Glycoproteins / Q000737:chemistry; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007251:Influenza, Human / Q000201:enzymology*; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry; D008958:Models, Molecular; D008968:Molecular Conformation; D009439:Neuraminidase / Q000737:chemistry* / Q000378:metabolism; D010957:Plasmids; D011487:Protein Conformation; D017434:Protein Structure, Tertiary
chemical_list: D006023:Glycoproteins; D008565:Membrane Proteins; D009439:Neuraminidase