THE H3/H4 HISTONE GENE CLUSTER OF LAND SNAILS (GASTROPODA: STYLOMMATOPHORA): TS/TV RATIO, GC3 DRIVE AND SIGNALS IN STYLOMMATOPHORAN PHYLOGENY

Histone gene primers were developed for land snails (Stylommatophora). The partial H3/H4 histone gene cluster was cloned and sequenced for 18 species. Transcription of the H3 and H4 genes was divergent (each gene is transcribed in the opposite direction) as has been found for other protostome and diploblast animals, with the exception of Mytilus. In the bivalve Mytilus transcription of both genes occurs in the same direction, i.e. land snails and bivalves seem to differ in their histone gene organization. The non-transcribed H3/H4 spacer varied in length between 279 and 691 basepairs. Nucleotide polymorphisms in this non-transcribed spacer might be of significance to study phylogenetics and systematics of closely related species and genera. As expected, the coding regions exhibited no amino acid substitution among land snail species. However, one amino acid substitution was found in comparison between land snails and Drosophila. The transition/transversion (TS/TV) ratio of H3 and H4 was predominately shaped by the third codon position and ranged in most cases from 1.0 to 2.0, indicating low nucleotide saturation. GC content was calculated for the third codon position (GC3 index at the ‘wobble' base position). The histone GC3 values were far lower in land snails than values currently available for other genomes (i.e. mammals). This indicates that H3/H4 histone wobble bases of land snails evolve without strong GC drive. Phylogenetic trees were reconstructed from the coding regions. We used Succinea putris (Elasmognatha) as outgroup. Trichia villosa (Helicoidea) showed six apomorphic nucleotide signals. Moreover, the nucleotide signals give evidence that the Cochlicopidae, Vertiginidae and Valloniidae are paraphyletic family categories. The paraphyletic status of cochlicopid, vertiginid and valloniid gastropods is also supported by our unpublished ribosomal DNA tree