Microarray analysis of the in vivo sequence preferences of a minor groove binding drug.
BACKGROUND
Minor groove binding drugs (MGBDs) interact with DNA in a sequence-specific manner and can cause changes in gene expression at the level of transcription. They serve as valuable models for protein interactions with DNA and form an important class of antitumor, antiviral, antitrypanosomal and antibacterial drugs. There is a need to extend knowledge of the sequence requirements for MGBDs from in vitro DNA binding studies to living cells.


RESULTS
Here we describe the use of microarray analysis to discover yeast genes that are affected by treatment with the MGBD berenil, thereby allowing the investigation of its sequence requirements for binding in vivo. A novel approach to sequence analysis allowed us to address hypotheses about genes that were directly or indirectly affected by drug binding. The results show that the sequence features of A/T richness and heteropolymeric character discovered by in vitro berenil binding studies are found upstream of genes hypothesized to be directly affected by berenil but not upstream of those hypothesized to be indirectly affected or those shown to be unaffected.


CONCLUSIONS
The data support the conclusion that effects of berenil on gene expression in yeast cells can be explained by sequence patterns discovered by in vitro binding experiments. The results shed light on the sequence and structural rules by which berenil binds to DNA and affects the transcriptional regulation of genes and contribute generally to the development of MGBDs as tools for basic and applied research.
Eckdahl
Todd T
TT
Biology Department, Missouri Western State University, Saint Joseph, MO, 64507, USA. eckdahl@missouriwestern.edu
Brown
Adam D
AD
Hart
Steven N
SN
Malloy
Kelly J
KJ
Shott
Martha
M
Yiu
Gloria
G
Hoopes
Laura L Mays
LL
Heyer
Laurie J
LJ
D001665:Binding Sites; D002478:Cells, Cultured; D004133:Diminazene / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D007364:Intercalating Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D046228:Microarray Analysis; D009690:Nucleic Acid Conformation / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods*
D007364:Intercalating Agents; D012333:RNA, Messenger; C003915:diminazene aceturate; D004133:Diminazene