Structural deviations in a bovine low expression lysozyme-encoding gene active in tissues other than stomach

Abstract

Lysozyme-encoding genes (Lys) constitute a gene-family in ruminants. While several of these genes are highly expressed in stomach (sLys), few other copies are weakly expressed in other tissues, notably in polymorphnuclear granulocytes and macrophages ( mLys). Searching an understanding for these grossly different levels of expression, we isolated the bovine variant of the gene being expressed in granulocytes and characterized it by sequencing, together with its promoter. Spanning about 9 kb of genomic DNA, the gene is found to be segmented into four exons, in common with all other Lys, as known from vertebrates. Sequence homologies between all bovine sLys-variants exceeds 70% over much of the entire coding sequence and promoter region. This indicates (i) that bovine lysozymes expressed either in stomach or granulocyte originate from a common ancestral gene and (ii) also excludes the possibility that the observed weak expression of the mLys gene is due to major structural rearrangements within the promoter segment. However, primer extension analysis based on RNA isolated from kidney locates the transcription startpoint (tsp of that gene) 44 nt further upstream than observed in both, bovine stomach lysozyme RNA or any of the homologous genes in mice and man. The observed weak expression of this bovine mLys gene appears to be a consequence of both the presence of an extra ATG codon in the extended 5′-UTR, and a severe down mutation of the ancestral TATA-box which is only partially compensated for by the presence of another mutation further upstream resulting in a weak substitute promoter sequence.