Double-stranded DNA binding characteristics and subcellular distribution of a minor groove binding diphenyl ether bisbenzimidazole.
The interactions of Hoechst 33377 (H1) with 20 different oligomeric duplexes have been investigated via spectrofluorometric titrations and/or thermal denaturation experiments. H1 is shown to form 2:1 complexes with dsDNA binding sites of at least four contiguous A/T base pairs. H1 is also shown to possess the rare ability to meaningfully distinguish between different A.T rich sequences. For example, the combined equilibrium constants for complexation of the oligomeric duplex 5'-GCAATTGC-3' (15) by H1 are found to be 110-fold greater than for the duplex 5'-GCTTAAGC-3' (16). It is believed that the 5'-TpA-3' dinucleotide step in 16 disrupts the rigid "A-tract" conformation of 15 and discourages minor groove binding by agents capable of recognizing longer dsDNA sequences. Molecular models are presented which elucidate the structure of the (H1)(2)-dsDNA minor groove complex. The two H1 molecules bind to an A/T rich sequence of 6 bp in a slightly staggered, side-by-side, and antiparallel arrangement. Evidence suggests that the piperazine rings of the H1 side-by-side complex are capable of resting in the minor groove of G/C base pairs. Fluorescence microscopy studies using NIH3T3 cells indicate that H1 is capable of traversing the cytoplasmic membrane and selectively localizing to nuclear DNA. H1 also demonstrated the ability to inhibit endogenous transcription of the c-fos gene in NIH3T3 cells at micromolar concentrations. Cytotoxicity studies employing the same cell type show H1 to possess an LD(50) of 3.5 microM.
Satz
A L
AL
Department of Chemistry and Biochemistry, University of California at Santa Barbara, Santa Barbara, California 93106, USA.
White
C M
CM
Beerman
T A
TA
Bruice
T C
TC
D016475:3T3 Cells; D000818:Animals; D001562:Benzimidazoles / Q000138:chemical synthesis / Q000378:metabolism* / Q000633:toxicity; D001665:Binding Sites; D006690:Bisbenzimidazole / Q000378:metabolism / Q000633:toxicity; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D016762:Genes, fos / Q000187:drug effects; D007364:Intercalating Agents / Q000138:chemical synthesis / Q000378:metabolism* / Q000633:toxicity; D008024:Ligands; D051379:Mice; D008858:Microscopy, Phase-Contrast; D008958:Models, Molecular; D009692:Nucleic Acid Heteroduplexes / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D013050:Spectrometry, Fluorescence; D013347:Subcellular Fractions / Q000378:metabolism; D013816:Thermodynamics
D001562:Benzimidazoles; C432595:HOE 33377; D007364:Intercalating Agents; D008024:Ligands; D009692:Nucleic Acid Heteroduplexes; D012333:RNA, Messenger; D004247:DNA; D006690:Bisbenzimidazole; C017807:bisbenzimide ethoxide trihydrochloride