Sequence-specific minor groove binding ligands as potential regulators of gene expression in Xenopus laevis oocytes.
The mouse mammary tumor virus (MMTV) promoter is induced by glucocorticoid hormone. A robust hormone- and receptor-dependent gene activation could be reproduced in Xenopus laevis oocytes. The homogeneous response in this system allowed a detailed analysis of the DNA-protein interactions following hormone activation. The strategy of artificial regulating of gene activity by sequence-specific minor groove binding ligands is very attractive. We have synthesized and studied the interaction with DNA of bis-linked netropsin derivatives in which two monomers are attached via short linkers in head-to-head and tail-to-tail manners. We have found that cis-diammine-platinum bridged bis-netropsin added to Xenopus oocytes media penetrates cellular and nuclear membrane and binds selectively to the MMTV promoter at the DNA segment that partly overlaps with the site recognized by glucocorticoid receptor. DNase I footprinting studies demonstrate that there are more stronger binding sites for cis-diammine-platinum bridged bis-netropsin on the naked MMTV DNA which are found to be inaccessible for its binding in oocytes.
Belikov
S V
SV
Department of Cell and Molecular Biology, The Medical Nobel Institute, Karolinska Institute, SE 17177, Stockholm, Sweden.
Grokhovsky
S L
SL
Isaguliants
M G
MG
Surovaya
A N
AN
Gursky
G V
GV
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D008024:Ligands; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D008969:Molecular Sequence Data; D009429:Netropsin / Q000031:analogs & derivatives* / Q000494:pharmacology; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012689:Sequence Homology, Nucleic Acid; D015533:Transcriptional Activation; D014982:Xenopus laevis / Q000378:metabolism
D008024:Ligands; D009429:Netropsin; C032161:bis-netropsin; D003850:Deoxyribonuclease I