A chromosome end satellite of Rhynchosciara americana (Diptera: Sciaridae) resembling nematoceran telomeric repeats

Abstract

The characterization of chromosome end sequences of Rhynchosciara americana was initiated with the screening of a plasmid microlibrary made from a microdissected polytene chromosome tip. A 268 bp insert chosen for analysis hybridized specifically to non-telocentric chromosome ends in which reverse transcriptase had been previously identified. Southern-blot hybridization of R. americana genomic DNA cut with XbaI, the only restriction site identified in the 268 bp probe, showed a ladder composed of multimers of a band in the range of 400 bp indicating a tandem array of the repeat partially represented in the cloned fragment. The complete repeat unit obtained by inverse PCR is 414 bp long, 67% AT-rich, characterized by the restriction sites XbaI and SalI and displays features typical of a nematoceran telomeric satellite. The telomere-like repeat is apparently absent from the chromosomes of two other Rhynchosciara species, R. baschanti and R. milleri. Double staining for satellite hybridization and reverse transcriptase in R. americana suggests that the arrays composed of the telomeric-like satellite do not reach the chromosome ends. This may indicate that telomeric-like features of some nematoceran terminal satellites do not warrant their telomeric position.