Structure, biological activity of the upstream regulatory sequence, and conserved domains of a middle molecular mass neurofilament gene of Xenopus laevis.
During development, the molecular compositions of neurofilaments (NFs) undergo progressive modifications that correlate with successive stages of axonal outgrowth. Because NFs are the most abundant component of the axonal cytoskeleton, understanding how these modifications are regulated is essential for knowing how axons control their structural properties during growth. In vertebrates ranging from lamprey to mammal, orthologs of the middle molecular mass NF protein (NF-M) share similar patterns of expression during axonal outgrowth, which suggests that these NF-M genes may share conserved regulatory elements. These elements might be identified by comparing the sequences and activities of regulatory domains among the vertebrate NF-M genes. The frog, Xenopus laevis, is a good choice for such studies, because its early neural development can be observed readily and because transgenic embryos can be made easily. To begin such studies, we isolated genomic clones of Xenopus NF-M(2), tested the activity of its upstream regulatory sequence (URS) in transgenic embryos, and then compared sequences of regulatory regions among vertebrate NF-M genes to search for conserved elements. Studies with reporter genes in transgenic embryos found that the 1. 5 kb URS lacked the elements sufficient for neuron-specific gene expression but identified conserved regions with basal regulatory activity. These studies further demonstrated that the NF-M 1.5 kb URS was highly susceptible to positional effects, a property that may be relevant to the highly variant, tissue-specific expression that is seen among members of the intermediate filament gene family. Non-coding regions of vertebrate NF-M genes contained several conserved elements. The region of highest conservation fell within the 3' untranslated region, a region that has been shown to regulate expression of another NF gene, NF-L. Transgenic Xenopus may thus prove useful for testing further the activity of conserved elements during axonal development and regeneration.