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PMID: 1731330 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Discrimination between related DNA sites by a single amino acid residue of Myc-related basic-helix-loop-helix proteins.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 2 ·1992-01-15 ·Pages 599-602

Dang CV, Dolde C, Gillison ML, Kato GJ

Abstract

A yeast genetic system was developed to study how the basic regions of basic-helix-loop-helix (bHLH) proteins distinguish between related consensus bHLH binding sites, with nucleotide sequence CANNTG. The yeast bHLH protein CBF1 binds to the sequence CAC(A/G)TG found in the yeast centromere element CDE1 and in promoter regions of several yeast genes involved in methionine biosynthesis. Using a functional assay to rescue a mutant cbf1 yeast strain from methionine auxotrophy, we determined that the basic region of CBF1 could be replaced by the homologous region of either the vertebrate USF transcription factor or c-Myc, both of which bind CACGTG. The homologous region of the AP4 transcription factor, which recognizes the sequence CAGCTG, could not functionally replace the CBF1 basic region. However, only a single substitution, Met----Arg, in the AP4 basic region of the inactive chimera CBF-AP4 was sufficient to restore CBF1 function. In randomization experiments, only arginine or lysine provided functional substitutions at the AP4 methionine position. The results suggest that this conserved arginine residue in the basic regions of Myc-related bHLH proteins discriminates between CAC(A/G)TG and related sites.

Related Genes
CBF1
MeSH Terms
Amino Acid Sequence Arginine/chemistry Base Sequence Binding Sites DNA Mutational Analysis DNA-Binding Proteins/chemistry Molecular Sequence Data Proto-Oncogene Proteins c-myc/metabolism Saccharomyces cerevisiae/chemistry Structure-Activity Relationship Transcription Factors/chemistry
Chemicals
DNA-Binding Proteins Proto-Oncogene Proteins c-myc Transcription Factors Arginine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dang C V
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Dolde C
Gillison M L
Kato G J
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27 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.2.599
Published
1992-01-15
Pages
599-602
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC48286
Subset
IM
Grants
NCI NIH HHS · CA51497 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.2.599 PMC Full Text View journal details
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