PMID- 10419522
OWN - NLM
STAT- MEDLINE
DCOM- 19990819
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 31
DP  - 1999 Jul 30
TI  - Sequence selectivity of c-Myb in vivo. Resolution of a DNA target specificity 
      paradox.
PG  - 21986-94
AB  - We have investigated the basis for the striking difference between the broad DNA 
      sequence selectivity of the c-Myb transcription factor minimal DNA-binding domain 
      R(2)R(3) in vitro and the more restricted preference of a R(2)R(3)VP16 protein 
      for Myb-specific recognition elements (MREs) in a Saccharomyces cerevisiae 
      transactivation system. We show that sequence discrimination in yeast is highly 
      dependent on the expression level of Myb effector protein. Full-length c-Myb and 
      a C-terminally truncated protein (residues 1-360) were also included in the 
      study. All of the tested Myb proteins displayed very similar DNA binding 
      properties in electrophoretic mobility shift assays. Only minor differences 
      between full-length c-Myb and truncated c-Myb(1-360) were observed. In 
      transactivation studies in CV-1 cells, the MRE selectivity was highest at low 
      expression levels of Myb effector proteins. However, the discrimination between 
      MRE variants was rapidly lost with high input levels of effector plasmid. In 
      c-Myb-expressing K-562 cells, the high degree of MRE selectivity was retained, 
      thereby confirming the relevance of the results obtained in the yeast system. 
      These data suggest that the MRE selectivity of c-Myb is an intrinsic property of 
      only the R(2)R(3) domain itself and that the transactivation response of a 
      specific MRE in vivo may be highly dependent on the expression level of the Myb 
      protein in the cell.
FAU - Andersson, K B
AU  - Andersson KB
AD  - Department of Biochemistry, University of Oslo, N-0316 Oslo 3, Norway. 
      k.b.andersson@ioks.uio.no
FAU - Berge, T
AU  - Berge T
FAU - Matre, V
AU  - Matre V
FAU - Gabrielsen, O S
AU  - Gabrielsen OS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Oligodeoxyribonucleotides)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (Proto-Oncogene Proteins c-myb)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Trans-Activators)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Binding Sites
MH  - COS Cells
MH  - Cell Line
MH  - DNA-Binding Proteins/metabolism
MH  - Genes, Reporter
MH  - Humans
MH  - K562 Cells
MH  - Luciferases/genetics
MH  - Molecular Sequence Data
MH  - Oligodeoxyribonucleotides/chemistry/metabolism
MH  - *Promoter Regions, Genetic
MH  - Proto-Oncogene Proteins/genetics/*metabolism
MH  - Proto-Oncogene Proteins c-myb
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - Saccharomyces cerevisiae/genetics
MH  - Substrate Specificity
MH  - Trans-Activators/genetics/*metabolism
MH  - Transcriptional Activation
MH  - Transfection
EDAT- 1999/07/27 00:00
MHDA- 1999/07/27 00:01
CRDT- 1999/07/27 00:00
PHST- 1999/07/27 00:00 [pubmed]
PHST- 1999/07/27 00:01 [medline]
PHST- 1999/07/27 00:00 [entrez]
AID - S0021-9258(19)72475-2 [pii]
AID - 10.1074/jbc.274.31.21986 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Jul 30;274(31):21986-94. doi: 10.1074/jbc.274.31.21986.