Effects of nucleoside analog incorporation on DNA binding to the DNA binding domain of the GATA-1 erythroid transcription factor.
We investigate here the effects of the incorporation of the nucleoside analogs araC (1-beta-D-arabinofuranosylcytosine) and ganciclovir (9-[(1,3-dihydroxy-2-propoxy)methyl] guanine) into the DNA binding recognition sequence for the GATA-1 erythroid transcription factor. A 10-fold decrease in binding affinity was observed for the ganciclovir-substituted DNA complex in comparison to an unmodified DNA of the same sequence composition. AraC substitution did not result in any changes in binding affinity. 1H-15N HSQC and NOESY NMR experiments revealed a number of chemical shift changes in both DNA and protein in the ganciclovir-modified DNA-protein complex when compared to the unmodified DNA-protein complex. These changes in chemical shift and binding affinity suggest a change in the binding mode of the complex when ganciclovir is incorporated into the GATA DNA binding site.
Foti
M
M
Walt Disney Memorial Cancer Institute at Florida Hospital, Orlando 32826, USA.
Omichinski
J G
JG
Stahl
S
S
Maloney
D
D
West
J
J
Schweitzer
B I
BI
D000818:Animals; D000963:Antimetabolites / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D002645:Chickens; D003561:Cytarabine / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D050981:Erythroid-Specific DNA-Binding Factors; D015774:Ganciclovir / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009584:Nitrogen / Q000737:chemistry; D009705:Nucleosides / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011522:Protons; D020218:Response Elements / Q000235:genetics*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
D000963:Antimetabolites; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D009705:Nucleosides; D011522:Protons; D014157:Transcription Factors; D003561:Cytarabine; D004247:DNA; D009584:Nitrogen; D015774:Ganciclovir