Altered tissue specificity of the revertant shrunken allele upon Dissociation (Ds) excision is associated with loss of expression and molecular rearrangement at the corresponding non-allelic isozyme locus in maize

Summary Transposable element Activator (Ac) induced wild-type stable revertants, derived from McClintock's Dissociation (Ds) insertion shrunken (sh) mutant sh-m5933, have been examined for sucrose synthases, SS1 and SS2, encoded by the revertant (Sh) locus and the non-allelic gene Sus (previously designated as Ss2), respectively. A structurally normal Sh locus has been previously described in these revertants. Immuno-blot (Western) and Southern hybridization analyses reported here identify one of the nine alleles, Sh-r5, as unique for several features. It showed altered tissue specificity, as the SS1 protein encoded by the Sh-r5 allele was readily detectable in the immature embryo which is otherwise characterized by the Sus expression only. The level of Sh-r5 expression at the protein and enzyme level was marked by endosperm specific SS1 abundance and a significant down-regulation in the embryo similar to the standard Sh and Sus loci in endosperm and embryo, respectively. We infer that tissue specific levels of gene expression among maize Ss genes is significantly determined by trans-regulatory factors present in these two tissues. The Sh-r5 strain also exhibited a complete loss of the Sus expression in all tissues tested in the plant. Lack of any detectable phenotypic abnormality in the Sh-r5 strain due to the loss of SS2 protein indicated that either the SS2 protein is nonessential or that the two SS isozymes are functionally compensatory. Genomic filter hybridizations with the Sus cDNA clone indicated that the Sus locus in the Sh-r5 strain was not deleted and was, in fact, unique among these revertants. Together, these data provide an unusual insight into the regulation and function of the two SS isozymes in the maize plant.