Comparison between site-specific DNA binding proteins of male and female Schistosoma mansoni.
Several amplicons with approximately 120 bp each, obtained from the upstream domain of Schistosoma mansoni female-specific gene F-10, were coupled to Dynabeads M-280 streptavidin. The beads were used as a matrix for affinity purification of nuclear proteins obtained from mixed populations of adult worms. A protein of approximately 12 kDa, bound to the DNA in a sequence-independent manner. In contrast, when the DNA matrix was narrowed down to smaller synthetic oligonucleotides, bearing sequences corresponding to the TATA box and the CAAT box, band-shift assays revealed that different nuclear proteins from either adult male or female worms formed complexes with the DNA adduct. In order to characterise the bound proteins, the same oligonucleotides were UV cross-linked to the male and female protein extracts. Whilst the band shift experiments showed that the proteins from each sex produced a distinct mobility pattern when the TATA box sequences were tested and a similar one when the CAAT box sequences were added to the proteins, UV cross-linking experiments revealed clear qualitative differences between both, male and female proteins and also between the proteins binding to the two motifs. These results are compatible with a model in which the differential expression of the F-10 gene might depend on individual sub-sets of proteins.
Fantappié
M R
MR
Departamento de Bioquímica Médica ICB/CCS Universidade Federal do Rio de Janeiro, Brazil.
Correa-Oliveira
R
R
Caride
E C
EC
Geraldo
E A
EA
Agnew
A
A
Rumjanek
F D
FD
D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins / Q000235:genetics*; D004527:Egg Proteins / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D017238:Genes, Helminth / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism / Q000528:radiation effects; D009841:Oligonucleotides / Q000378:metabolism / Q000528:radiation effects; D009866:Oogenesis / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012550:Schistosoma mansoni / Q000737:chemistry / Q000235:genetics*; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; D004527:Egg Proteins; D015801:Helminth Proteins; D009687:Nuclear Proteins; D009841:Oligonucleotides; C084248:major eggshell protein F-10, Schistosoma mansoni