Antiparallel expression of the sense and antisense transcripts of maize alpha-tubulin genes

Abstract

In all eukaryotes alpha- and beta-tubulins are encoded by small families of closely related genes and are highly conserved. In Zea mays, at least six different alpha-tubulin coding sequences are known. We describe the isolation from scutellar nodes of the maize inbred line W22 of a clone (CTM5) coding for an alpha-tubulin. On the basis of the 3' end nucleotide sequence, this clone can be assigned to the already reported tua4 gene. Northern analysis demonstrates that CTM5 encodes a 1.5 kb transcript, which is expressed in different tissues of the seed and of the seedling. In order to define the spatial and temporal expression of alpha-tubulin genes, in situ hybridization experiments were performed on these tissues. Unexpectedly, a specific signal was detected with both antisense and sense RNA strands. Temporal and spatial distribution of the two RNAs, however, shows that high levels of the two transcripts are always discordant. In tissues where sense transcripts are highly abundant (embryos at various developmental stages, root tips, pollen grains), the antisense transcripts are expressed in relatively small amounts, while in pericarp, coleoptile, leaves, and scutellar node, where antisense transcripts accumulate, the sense transcript only reaches a very low level. Northern analysis using single-stranded DNA probes confirmed the presence of an antisense transcript of 1.5 kb, prompting speculation about the role of this transcript in the regulation of the expression of alpha-tubulin genes.