Proper spacing between heptad repeat B and the transmembrane domain boundary of the paramyxovirus SV5 F protein is critical for biological activity

Virology
J ZhouR A Lamb

Abstract

The paramyxovirus, simian virus 5, fusion (F) protein contains seven amino acids between heptad repeat B (a domain required for a biologically active fusion protein) and the presumptive boundary of the transmembrane (TM) domain. The role of the seven membrane proximal residues in stability and fusion promotion was examined by construction of a series of insertion, substitution, and deletion mutants, as manipulation of this region to enable proteolytic cleavage would facilitate production of a soluble F protein. The majority of the mutant F proteins both oligomerized and had kinetics of intracellular transport similar to those of wild-type (wt) F protein. All mutant F proteins were expressed at the cell surface at or near the same level as the wt F protein. However, by using both a qualitative lipid mixing assay and a quantitative content mixing assay for membrane fusion, it was found that mutant F proteins containing insertions in the region between heptad repeat B and the TM domain were unable to induce fusion, whereas the mutant F proteins containing substitutions in this region, together with three of the four mutants with deletions in this region, could induce fusion. Four of the F protein mutants contained a Factor Xa clea...Continue Reading

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Citations

Sep 25, 1999·Journal of Molecular Biology·B EisenhaberF Eisenhaber
Apr 12, 2002·Trends in Biochemical Sciences·Sergio G Peisajovich, Yechiel Shai
Apr 13, 1999·Molecular Cell·K A BakerT S Jardetzky
Jan 6, 2009·Journal of Molecular Biology·Mei Lin Z BissonnetteRobert A Lamb
Apr 22, 2008·Biochimica Et Biophysica Acta·Ana J Pérez-BernáJosé Villalaín
Nov 5, 2003·Nature Structural Biology·Heather E ParkJudith M White

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