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Analysis of the Cytoplasmic Tail Domain of Hendra Virus Fusion Protein
Author(s) -
Gibson Kathleen Marie,
Wurth Mark Allen,
Dutch Rebecca Ellis
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a966
Subject(s) - fusion protein , lipid bilayer fusion , cytoplasm , biology , glycoprotein , cell fusion , protein domain , microbiology and biotechnology , mutant , hendra virus , transmembrane domain , transmembrane protein , biochemistry , virus , amino acid , cell , virology , gene , ebola virus , recombinant dna , receptor
Hendra virus (HeV), a recently emerged paramyxovirus, produces two glycoproteins: a fusion (F) protein and an attachment protein, which together mediate virus‐cell fusion and cell‐cell fusion. The F protein is produced in a precursor form, F 0 , which must be proteolytically cleaved into the disulfide‐linked F 1 and F 2 heterodimer by the endosomal protease cathepsin L to be fusogenically active. The Hendra F protein also contains four N‐linked carbohydrate additions, two of which have been shown to affect protein folding and membrane fusion. The F 1 portion of the fusion protein has a number of important domains, including a fusion peptide, two heptad repeat regions, a transmembrane domain and 28 amino acid cytoplasmic tail domain. While mutations in the cytoplasmic tail regions have been studied for a number of paramyxovirus F proteins, the role of this domain in folding, processing and membrane fusion is still unclear. We have created a series of deletion mutants along the cytoplasmic tail domain of the Hendra F protein, resulting in proteins with cytoplasmic tails of varying length. All of the deletion mutants were expressed in transient cell culture expression system. However, heterogeneity in size was observed for a number of mutants, and this could be resolved by removal of the N‐linked carbohydrates, suggesting that the cytoplasmic tail can affect branching of the N‐linked carbohydrates. In addition, alterations in fusion properties were observed for some of the mutants, indicating that this cytoplasmic domain also plays a role in membrane fusion promotion of this important viral protein. Funded by grants from NIH.