A genomic change associated with the development of resistance to hycanthone in Schistosoma mansoni.
Ribosomal gene probes were used to investigate the genetic basis of drug resistance in schistosomes in a model where resistance to the anthelmintic hycanthone (HC) is generated by exposing immature worms to the drug. Two strains of Schistosoma mansoni, JHU and NMRI, were used. Drug resistance could be produced in the JHU strain by treatment with HC, but was also found to occur spontaneously. In contrast, it was not possible to detect or produce resistance to HC in the NMRI strain. A genomic alteration accompanied the development of resistance. The change was evidence by the occurrence of restriction fragment length polymorphisms (RFLPs) when Southern blots of genomic DNA from HC-resistant worms were hybridized with the ribosomal probe pSM389, which contains part of the small rRNA gene plus non-transcribed spacer (NTS) sequence. The most reliable marker of HC-resistance was a 3.6-kb BamHI fragment which was present and heritable in 7 drug-resistant lines derived from the JHU strain but absent from the parent JHU population and from NMRI parasites. The universal absence of the 3.6-kb RFLP in HC-sensitive individuals and its presence in the drug-resistant progeny suggest that resistance results from an induced change in the population rather than from selection of HC-resistant parasites. The rRNA gene sequence responsible for detecting the 3.6-kb RFLP appears to be localized either to the NTS or to the 5' end of the small rRNA gene, since hybridization to a probe containing sequence from the rRNA gene contiguous and downstream from the insert of pSM389 failed to reveal the RFLP. These results show that the development of resistance to HC is accompanied by a genomic rearrangement.
Brindley
P J
PJ
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Lewis
F A
FA
McCutchan
T F
TF
Bueding
E
E
Sher
A
A
D000818:Animals; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis / Q000302:isolation & purification; D004351:Drug Resistance / Q000235:genetics*; D006826:Hycanthone / Q000494:pharmacology*; D051379:Mice; D010957:Plasmids; D012150:Polymorphism, Restriction Fragment Length; D015347:RNA Probes; D012335:RNA, Ribosomal / Q000235:genetics; D012550:Schistosoma mansoni / Q000187:drug effects / Q000235:genetics*; D013892:Thioxanthenes / Q000494:pharmacology*
D015347:RNA Probes; D012335:RNA, Ribosomal; D013892:Thioxanthenes; D006826:Hycanthone; D004247:DNA