Document Detail


An E-box-mediated increase in cad transcription at the G1/S-phase boundary is suppressed by inhibitory c-Myc mutants.
MedLine Citation:
PMID:  7739536     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To better understand the signaling pathways which lead to DNA synthesis in mammalian cells, we have studied the transcriptional activation of genes needed during the S phase of the cell cycle. Transcription of the gene encoding a pyrimidine biosynthetic enzyme, carbamoyl-phosphate synthase (glutamine-hydrolyzing)/aspartate carbamoyltransferase/dihydroorotase (cad), increases at the G1/S-phase boundary. We have mapped the growth-dependent response element in the hamster cad gene to the extended palindromic E-box sequence, CCACGTGG, which is centered at +65 in the 5' untranslated sequence. Mutation of the E box abolished growth-dependent transcription, and an oligonucleotide corresponding to the cad sequence at +55 to +75 (+55/+75) restored growth-dependent regulation to nonresponsive cad promoter mutants when placed down-stream of the transcription start site. The same oligonucleotide conferred less G1/S-phase induction when placed upstream of basal promoter elements. An analogous oligonucleotide containing the mutant E box had no effect in either location. Nuclear proteins bound the cad +55/+75 element in a cell cycle-dependent manner in electromobility shift assays; antibodies specific to USF and Max blocked the DNA-binding activity of different growth-regulated protein-DNA complexes. Expression of c-Myc mutants which have been shown to dominantly interfere with the function of c-Myc and Max significantly inhibited cad transcription during S phase but had no effect on transcription from another G1/S-phase-activated promoter, dhfr. These data support a model whereby E-box-binding proteins activate serum-induced transcription from the cad promoter at the G1/S-phase boundary and suggest that a Max-associated protein complex contributes to the serum response.
Authors:
R J Miltenberger; K A Sukow; P J Farnham
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Molecular and cellular biology     Volume:  15     ISSN:  0270-7306     ISO Abbreviation:  Mol. Cell. Biol.     Publication Date:  1995 May 
Date Detail:
Created Date:  1995-06-02     Completed Date:  1995-06-02     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  8109087     Medline TA:  Mol Cell Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2527-35     Citation Subset:  IM    
Affiliation:
McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School 53706, USA.
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MeSH Terms
Descriptor/Qualifier:
3T3 Cells
Animals
Aspartate Carbamoyltransferase / genetics*
Base Sequence
Binding Sites / genetics
Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / genetics*
Cricetinae
DNA / genetics,  metabolism
Dihydroorotase / genetics*
G1 Phase / genetics
Genes, myc*
Mesocricetus
Mice
Molecular Sequence Data
Multienzyme Complexes / genetics*
Mutation
Promoter Regions, Genetic
S Phase / genetics
Suppression, Genetic
Transcription, Genetic
Grant Support
ID/Acronym/Agency:
CA07175/CA/NCI NIH HHS; CA09135/CA/NCI NIH HHS; CA59524/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/CAD trifunctional enzyme; 0/Multienzyme Complexes; 9007-49-2/DNA; EC 2.1.3.2/Aspartate Carbamoyltransferase; EC 3.5.2.3/Dihydroorotase; EC 6.3.5.5/Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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