DNA-binding domain of human c-Myc produced in escherichia coli

Journal Article · Thu Jun 01 04:00:00 EDT 1989 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.9.6.2477· OSTI ID:5604401

Dang, C V; Buckmire, M; VanDam, H; Lee, W M.F.

The authors have identified the domain of the human c-myc protein (c-Myc) produced in Escherichia coli that is responsible for the ability of the protein to bind sequence-nonspecific DNA. Using analysis of binding of DNA by proteins transferred to nitrocellulose, DNA-cellulose chromatography, and a nitrocellulose filter binding assay, they examined the binding properties of c-Myc peptides generated by cyanogen bromide cleavage, of butane c-,Myc, and of proteins that fuse portions of c-Myc to staphylococcal protein A. The results of these analyses indicated that c-Myc amino acid 265 to 318 were responsible for DNA binding and that other regions of the protein (including a highly conserved basic region and a region containing the leucine zipper motif) were not required. Some mutant c-Mycs that did not bind DNA maintained rat embryo cell-cotransforming activity, which indicated that the c-Myc property of in vitro DNA binding was not essential for this activity. These mutants, however, were unable to transform established rat fibroblasts (Rat-1a cells) that were susceptible to transformation by wild-type c-Myc, although this lack of activity may not have been due to their inability to bind DNA.

• Research Organization: Johns Hopkins Univ., Baltimore, MD (USA). School of Medicine; Medical Biochemistry, Sylvius Labs., 2300 RA Leiden (NL); Pennsylvania Univ., Philadelphia, PA (USA). School of Medicine
• OSTI ID: 5604401
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 9:6; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
ANIMAL CELLS
ANIMALS
BACTERIA
CHEMICAL BONDS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DNA
DNA SEQUENCING
EMBRYONIC CELLS
ESCHERICHIA COLI
FIBROBLASTS
GENES
MAMMALS
MICROORGANISMS
MOLECULAR STRUCTURE
MUTANTS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PEPTIDES
PROTEINS
RATS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES