PMID- 2717405
OWN - NLM
STAT- MEDLINE
DCOM- 19890614
LR  - 20190501
IS  - 0305-1048 (Print)
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Linking)
VI  - 17
IP  - 7
DP  - 1989 Apr 11
TI  - Measurement of the binding of transcription factor Sp1 to a single GC box 
      recognition sequence.
PG  - 2639-53
AB  - The equilibrium constant was determined for the binding of the transcription 
      factor Sp1 to a single consensus GC box DNA recognition site, (5'-GGGGCGGGGC-3'). 
      For these experiments, single copies of the recognition site were synthesized and 
      cloned in a standard plasmid background. Binding was measured either by a 
      footprinting assay modified so that the binding reaction was at equilibrium, or 
      by a gel mobility shift assay. The concentration of active Sp1 in the reactions 
      and the dissociation constant were determined by computer-assisted fitting to 
      theoretical curves. Values for the dissociation constant obtained in different 
      experiments ranged from 4.1 X 10(-10) M to 5.3 X 10(-10) M. Several variants of 
      the consensus recognition site were also tested. An A-substituted variant 
      (5'-GGGGAGGGGC-3') and a T-substituted variant (5'-GGGGTGGGGC-3') were bound 
      3-fold and 6-fold more weakly than the consensus site, respectively. A 
      G-substituted variant (5'-GGGGGGGGGC-3') was bound at least 30-fold more weakly 
      than the consensus site. These findings help distinguish between alternative 
      models for Sp1-DNA recognition. They are consistent with the presence of specific 
      hydrogen-bond contacts between Sp1 and the central C-G base pair, but provide no 
      particular evidence to support a model where local DNA structure is the dominant 
      factor in the interaction.
FAU - Letovsky, J
AU  - Letovsky J
AD  - Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
FAU - Dynan, W S
AU  - Dynan WS
LA  - eng
GR  - GM 35866/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Oligonucleotides)
RN  - 0 (Transcription Factors)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Base Composition
MH  - Base Sequence
MH  - Binding Sites
MH  - DNA/*metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Genetic Variation
MH  - HeLa Cells
MH  - Humans
MH  - Kinetics
MH  - Nucleotide Mapping
MH  - Oligonucleotides/*metabolism
MH  - Transcription Factors/*metabolism
PMC - PMC317648
EDAT- 1989/04/11 00:00
MHDA- 1989/04/11 00:01
CRDT- 1989/04/11 00:00
PHST- 1989/04/11 00:00 [pubmed]
PHST- 1989/04/11 00:01 [medline]
PHST- 1989/04/11 00:00 [entrez]
AID - 10.1093/nar/17.7.2639 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1989 Apr 11;17(7):2639-53. doi: 10.1093/nar/17.7.2639.