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

Journal Article · · Mol. Cell. Biol.; (United States)
DOI:https://doi.org/10.1128/MCB.9.6.2477· OSTI ID:5604401

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), Vol. 9:6
Country of Publication:
United States
Language:
English