Existence of xenobiotic response element binding in Dictyostelium.
Xenobiotic response element (XRE) is a core nucleotide sequence at the upstream of inducible target genes for the transcription factor aryl hydrocarbon receptor (AhR) that is responsible for recognition of exogenous environmental pollutants in eukaryotic cells. Gel retardation electrophoresis revealed the presence of binding of a radiolabeled probe containing XRE in both cytosolic and nuclear preparations of the slime mold Dictyostelium. Unlabeled XRE probe was more potent in competing for XRE binding in both fractions than unlabeled XRE probe with a point mutation at the core element. Limited proteolysis by V8 protease did not markedly affect XRE binding in both fractions, while XRE binding decreased during in vitro incubation at 30 degrees C for up to 24 h at decline rates proportional to increasing pHs at a range of 6.5-8.5 in cytosolic fractions in a manner different from those in nuclear fractions. Deprivation of nutrients induced aggregation of cells within 4-8 h later, followed by formation of first finger tips around 12 h later and subsequent development to mobile slugs within 16 h and then to fruit bodies between 20 and 24 h later. The starvation led to a marked decrement of XRE binding in cytosolic fractions 4-36 h later, followed by a robust but transient increment of that in nuclear extracts 12-20 h afterward. However, XRE binding was not affected by antibodies against AhR-related proteins known to date in both fractions irrespective of nutritional conditions. These results suggest the abundance of as-yet unidentified proteins with high affinity for XRE in the slime mold Dictyostelium. The possibility that those proteins may be translocated from the cytoplasm to the nucleus in response to cellular development during starvation is feasible.
Kuramoto
Nobuyuki
N
Department of Molecular Pharmacology, Kanazawa University Faculty of Pharmaceutical Sciences, 13-1 Takara-machi, Ishikawa 920-0934, Kanazawa, Japan. kuramoto@mail.p.kanazawa-u.ac.jp
Goto
Emi
E
Masamune
Yukito
Y
Gion
Keiko
K
Yoneda
Yukio
Y
D000818:Animals; D001345:Autoradiography; D002467:Cell Nucleus / Q000737:chemistry; D003600:Cytosol / Q000737:chemistry; D004023:Dictyostelium / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D016681:Genome, Fungal; D015151:Immunoblotting; D018336:Receptors, Aryl Hydrocarbon / Q000032:analysis; D020218:Response Elements; D014466:Ultraviolet Rays; D015262:Xenobiotics / Q000032:analysis*
D018336:Receptors, Aryl Hydrocarbon; D015262:Xenobiotics