Regulation of gene expression by sequence-specific alkylating polyamide.
In order to investigate the inhibition of gene expression by a new type of hairpin polyamide-CPI conjugate 1, its ability to inhibit transcription in cell free system was investigated. Sequence-selective alkylation of double-stranded DNA by 1 was investigated by denaturing gel electrophoresis using 1000 bp DNA fragment which codes for green fluorescence protein (GFP) under the control of T7 promoter. Analysis of DNA sequence indicated that 1 alkylated predominantly at the site of 5'-AGTCA-3' in coding region of GFP. The transcript by T7 RNA polymerase using the alkylated DNA as a template was analyzed by PAGE. The results clearly indicate that 1 inhibits transcription by alkylation of coding region at a nanomolar concentration.
Oyoshi
Takanori
T
Division of Biofunctional Molecules, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Surugadai, Kanda, Chiyoda, Tokyo 101-0062, Japan.
Kawakami
Wakana
W
Bando
Toshikazu
T
Narita
Akihiko
A
Sugiyama
Hiroshi
H
D000478:Alkylation; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000187:drug effects*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D009757:Nylons / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D008164:Luminescent Proteins; D009757:Nylons; D049452:Green Fluorescent Proteins