PMID- 8264603
OWN - NLM
STAT- MEDLINE
DCOM- 19940121
LR  - 20210526
IS  - 0270-7306 (Print)
IS  - 1098-5549 (Electronic)
IS  - 0270-7306 (Linking)
VI  - 14
IP  - 1
DP  - 1994 Jan
TI  - Dominant negative retinoid X receptor beta inhibits retinoic acid-responsive gene 
      regulation in embryonal carcinoma cells.
PG  - 360-72
AB  - Retinoid X receptors (RXRs) heterodimerize with multiple nuclear hormone 
      receptors and are thought to exert pleiotropic functions. To address the role of 
      RXRs in retinoic acid- (RA) mediated gene regulation, we designed a dominant 
      negative RXR beta. This mutated receptor, termed DBD-, lacked the DNA binding 
      domain but retained the ability to dimerize with partner receptors, resulting in 
      formation of nonfunctional dimers. DBD- was transfected into P19 murine embryonal 
      carcinoma (EC) cells, in which reporters containing the RA-responsive elements 
      (RAREs) were activated by RA through the activity of endogenous RXR-RA receptor 
      (RAR) heterodimers. We found that DBD- had a dominant negative activity on the 
      RARE reporter activity in these cells. P19 clones stably expressing DBD- were 
      established; these clones also failed to activate RARE-driven reporters in 
      response to RA. Further, these cells were defective in RA-induced mRNA expression 
      of Hox-1.3 and RAR beta, as well as in RA-induced down-regulation of Oct3 mRNA. 
      Gel mobility shift assays demonstrated that RA treatment of control P19 cells 
      induces RARE-binding activity, of which RXR beta is a major component. However, 
      the RA-induced binding activity was greatly reduced in cells expressing DBD-. By 
      genomic footprinting, we show that RA treatment induces in vivo occupancy of the 
      RARE in the endogenous RAR beta gene in control P19 cells but that this occupancy 
      is not observed with the DBD- cells. These data provide evidence that the 
      dominant negative activity of DBD- is caused by the lack of receptor binding to 
      target DNA. Finally, we show that in F9 EC cells expression of DBD- leads to 
      inhibition of the growth arrest that accompanies RA-induced differentiation. 
      Taken together, these results demonstrate that RXR beta and partner receptors 
      play a central role in RA-mediated gene regulation and in the control of growth 
      and differentiation in EC cells.
FAU - Minucci, S
AU  - Minucci S
AD  - Laboratory of Molecular Growth Regulation, National Institute of Child Health and 
      Human Development, National Institutes of Health, Bethesda, Maryland 20892.
FAU - Zand, D J
AU  - Zand DJ
FAU - Dey, A
AU  - Dey A
FAU - Marks, M S
AU  - Marks MS
FAU - Nagata, T
AU  - Nagata T
FAU - Grippo, J F
AU  - Grippo JF
FAU - Ozato, K
AU  - Ozato K
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Neoplasm)
RN  - 0 (Receptors, Cytoplasmic and Nuclear)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Retinoid X Receptors)
RN  - 0 (Transcription Factors)
RN  - 5688UTC01R (Tretinoin)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Carcinoma, Embryonal/genetics/metabolism/pathology
MH  - Cell Differentiation/genetics
MH  - Cell Division/genetics
MH  - DNA Primers/genetics
MH  - DNA, Neoplasm/genetics
MH  - Gene Expression Regulation, Neoplastic/*drug effects
MH  - Genes, Reporter
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Receptors, Cytoplasmic and Nuclear/*genetics/metabolism
MH  - Receptors, Retinoic Acid/genetics/metabolism
MH  - Retinoid X Receptors
MH  - *Transcription Factors
MH  - Transcription, Genetic
MH  - Transfection
MH  - Tretinoin/metabolism/*pharmacology
MH  - Tumor Cells, Cultured/metabolism/pathology
PMC - PMC358385
EDAT- 1994/01/01 00:00
MHDA- 1994/01/01 00:01
PMCR- 1994/01/01
CRDT- 1994/01/01 00:00
PHST- 1994/01/01 00:00 [pubmed]
PHST- 1994/01/01 00:01 [medline]
PHST- 1994/01/01 00:00 [entrez]
PHST- 1994/01/01 00:00 [pmc-release]
AID - 10.1128/mcb.14.1.360-372.1994 [doi]
PST - ppublish
SO  - Mol Cell Biol. 1994 Jan;14(1):360-72. doi: 10.1128/mcb.14.1.360-372.1994.