Differences in DNA sequence recognition by the heat-shock factors of Drosophila melanogaster and the parasitic helminth Schistosoma mansoni.
It was recently shown that schistosome extracts contain heat-shock factor (HSF) activity that correlates with the pattern of hsp70 mRNA levels at different developmental stages of the parasite (Levy-Holtzman and Schechter (1994) Parasitology 108, 35-42). To extend our understanding of the HSF activity revealed in extracts of Schistosoma mansoni (Sm), it was further analyzed by competition experiments and compared with the well characterized HSF of Drosophila melanogaster (Dm). The interactions of HSF in Sm extracts (SmHSF) and HSF of Dm (DmHSF) with 32P-labeled heat shock element (HSE) probes, with and without unlabeled competitor DNA probes (HSE-related oligos), were analyzed by gel retardation assay. The binding and inhibition studies demonstrated that SmHSF and DmHSF differ in HSE sequence recognition: an array of three nGAAn inverted repeats according to the ideal consensus sequence (nGAAnnTTCnnGAAn) is recognized by DmHSF, but not by SmHSF. In the schistosome, binding is attained only when the third pentamer is a variant, composed of nGTAn instead of nGAAn. The presence of this variant in the promoter of the hsp70 gene of the parasite suggests coevolution of the variant sequence together with the SmHSF which interacts efficiently with the variant, but not with the ideal HSE sequence. Further inhibition studies revealed additional differences between SmHSF and DmHSF in recognition of the first and second nGAAn pentamers of HSE. In analogy to other systems of ligand-protein interactions, we propose that the complementarity between the HSE ligand and the HSF protein is higher in SmHSF, as compared to DmHSF.
Levy-Holtzman
R
R
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Clos
J
J
Schechter
I
I
D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D015342:DNA Probes; D004331:Drosophila melanogaster / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012550:Schistosoma mansoni / Q000235:genetics*
D015342:DNA Probes; D006360:Heat-Shock Proteins; D004247:DNA