myc-related proteins and DNA sequences in Trypanosoma brucei

The cAMP content of Trypanosoma brucei increases in parallel with ascending mammalian parasitemia to very high levels just before differentiation of the long-slender to the short-stumpy bloodstream form. Because expression of myc oncogenes is required for vertebrate cells to interpret proliferation signals and declines in response to cAMP mediated differentiation, we investigated whether T. brucei also harbored myc-like proteins and genes. Accordingly, we probed lysates of long-slenders, short-stumpies and procyclics (insect midgut stage) with antibody to myc proteins and also hybridized myc gene family sequences to procyclic DNA. We found that antibody to myc-family proteins of mammals reacts with 40 kDa and 55 kDa proteins in all three life cycle stages, and that procyclic DNA contains three EcoRI fragments that are homologous to a v-myc probe. One of these fragments also hybridizes to a synthetic 25-mer oligonucleotide deduced from a consensus sequence in the second exon of the myc family and expresses a 3.2 kb mRNA transcript in Northern blots of procyclic RNA. The conservation of myc-family homologous across the broad phylogenetic gap between mammals and trypanosomes illustrates ancient evolutionary relationships and raises the possibility of stage-specific expression of myc genes during the life cycle of T. brucei.