C-terminal trimerization, but not N-terminal trimerization, of the reovirus cell attachment protein Is a posttranslational and Hsp70/ATP-dependent process.
The C-terminal globular head of the lollipop-shaped final sigma1 protein of reovirus is responsible for interaction with the host cell receptor. Like the N-terminal fibrous tail, it has its own trimerization domain. Whereas N-terminal trimerization (formation of a triple alpha-helical coiled coil) occurs at the level of polysomes (i.e. cotranslationally) and is ATP-independent, C-terminal trimerization is a posttranslational event that requires ATP. Coprecipitation experiments using anti-Hsp70 antibodies and truncated final sigma1 proteins synthesized in vitro revealed that only regions downstream of the N-terminal alpha-helical coiled coil were associated with Hsp70. Hsp70 was also found to be associated with nascent final sigma1 chains on polysomes as well as with immature postribosomal final sigma1 trimers (hydra-like intermediates with assembled N termini and unassembled C termini). These latter structures were true intermediates in the final sigma1 biogenetic pathway since they could be chased into mature final sigma1 trimers with the release of Hsp70. Thus, unlike N-terminal trimerization, C-terminal trimerization is Hsp70- and ATP-dependent. The involvement of two mechanistically distinct oligomerization events for the same molecule, one cotranslational and one posttranslational, may represent a common approach to the generation of oligomeric proteins in the cytosol.
Leone
G
G
Department of Microbiology and Infectious Diseases, University of Calgary, Health Sciences Centre, Calgary, Alberta, Canada T2N 4N1.
Coffey
M C
MC
Gilmore
R
R
Duncan
R
R
Maybaum
L
L
Lee
P W
PW
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D036022:Capsid Proteins; D002474:Cell-Free System; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D046911:Macromolecular Substances; D018832:Molecular Chaperones / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D011991:Receptors, Virus / Q000378:metabolism; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*
D036022:Capsid Proteins; D018840:HSP70 Heat-Shock Proteins; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D011991:Receptors, Virus; D014764:Viral Proteins; C031728:sigma 1 protein, reovirus; D000255:Adenosine Triphosphate