Assembly of adenovirus type 2 fiber synthesized in cell-free translation system.
Physicochemical and functional analyses of the translation products of fiber mRNA in rabbit reticulocyte lysate suggested that fiber polypeptide chains (monomers) were capable of self-assembling in vitro, forming trimeric fibers (trimers) without direct intervention of any other adenovirus-coded protein or cell nuclear matrix component. Kinetic studies showed that trimer formation occurred at a rate six times lower than that of fiber polypeptide synthesis. Fiber assembly was found to be relatively inefficient in vitro, with only 25-30% fiber polypeptides trimerized after 4-h translation reaction. The rate constant for fiber subunit assembly, extrapolated from the kinetic curves of trimer formation, was found to be in the order of magnitude of 10(5) M-1 s-1, with a t 1/2 of 1.3 h at 30 degrees C. A latence phase of approximately 40 min in the appearance of the first detectable trimers indicated that fiber assembly did not occur co-translationally, suggesting the existence of rate-limiting intermediate step(s) during assembly.
Novelli
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A
Laboratoire de Virologie et Pathogénèse Moléculaires, Faculté de Médecine, Montpellier, France.
Boulanger
P A
PA
D000260:Adenoviruses, Human / Q000648:ultrastructure*; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D009024:Morphogenesis; D014176:Protein Biosynthesis; D014357:Trypsin / Q000494:pharmacology; D014461:Ultracentrifugation; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
D036022:Capsid Proteins; D046911:Macromolecular Substances; D014764:Viral Proteins; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus; D014357:Trypsin