Abstract
The oligomerization domain of the reovirus cell attachment protein (sigma 1) was probed using the type 3 reovirus sigma 1 synthesized in vitro. Trypsin cleaved the sigma 1 protein (49K molecular weight) approximately in the middle and yielded a 26K N-terminal fragment and a 23K C-terminal fragment. Under conditions which allowed for the identification of intact sigma 1 in the oligomeric form (approximately 200K) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the N-terminal 26K fragment was found to exist as stable trimers (80K) and, to a less extent, as dimers (54K), whereas the C-terminal fragment remained in the monomeric form. A polypeptide (161 amino acids) containing the N-terminal heptad repeat region synthesized in vitro was capable of forming stable dimers and trimers. Using various criteria, we demonstrated that the stability of the intact sigma 1 oligomer is conferred mainly by the N-terminal heptad repeat region. Our results are summarized in a model in which individual heptad repeats are held together in a three-stranded alpha-helical coiled-coil structure via both hydrophobic and electrostatic interactions.
MeSH Terms
Amino Acid Sequence
Base Sequence
Capsid Proteins
Cloning, Molecular
Dithiothreitol
Ions
Macromolecular Substances
Mercaptoethanol
Molecular Sequence Data
Molecular Weight
Oligonucleotides/chemistry
Protein Binding
Reoviridae/analysis
Structure-Activity Relationship
Trypsin/pharmacology
Viral Proteins/chemistry,genetics
Chemicals
Capsid Proteins
Ions
Macromolecular Substances
Oligonucleotides
Viral Proteins
sigma 1 protein, reovirus
Mercaptoethanol
Trypsin
Dithiothreitol
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Leone G
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Alberta, Canada.
Duncan R
Mah D C
Price A
Cashdollar L W
Lee P W
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