MUTATIONS IN A CCHC ZINC-BINDING MOTIF OF THE REOVIRUS SIGMA-3 PROTEIN DECREASE ITS INTRACELLULAR STABILITY

It has been demonstrated that the sigma 3 protein of reovirus harbors a zinc-binding domain in its aminoterminal portion. A putative zinc finger in the CCHH form is located in this domain and was considered to be a good candidate for the zinc-binding motif. We performed site-directed mutagenesis to substitute amino acids in this region and demonstrated that many of these mutants, although expressed in COS cells, were unstable compared with the wild-type protein. Further analysis revealed that zinc-binding capability, as measured by retention on a zinc chelate affinity adsorbent, correlates with stability. These studies also allowed us to identify a CCHC box as the most probable zinc-binding motif.