This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Watt, R A Articles by Rosenberg, M Search for Related Content PubMed PubMed Citation Articles by Watt, R A Articles by Rosenberg, M

 Previous Article  |  Next Article 

Mol Cell Biol. 1985 March; 5(3): 448-456

Expression and characterization of the human c-myc DNA-binding protein.

ABSTRACT

In an effort to study in detail the nature of the protein product of the human protooncogene c-myc, we have expressed the gene at high levels in Escherichia coli. The c-myc coding region was taken from a full-length cDNA clone and inserted into a vector designed to express foreign gene products efficiently in E. coli. Pulse-labeling experiments indicated that the rate of expression of c-myc in this thermoinducible expression system is very efficient. The product was relatively stable and accumulated to approximately 10% of total cellular protein. A purification protocol was devised which allowed the c-myc protein to be readily purified in quantities sufficient for detailed biochemical and physical analyses. A high-titer polyclonal antiserum was raised against the pure protein and shown to immunoprecipitate the p110gag-myc fusion protein of MC-29-infected quail cells. This antiserum also selectively detects a protein with an apparent molecular weight of 64,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis from a Burkitt lymphoma cell line. We conclude that this 64-kilodalton protein is the human c-myc gene product since the E. coli-made protein exhibits an equivalent molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, even though its calculated molecular weight is 49,000. Furthermore, we demonstrate that the bacterially made human c-myc protein is a DNA-binding protein and that it exhibits a high nonspecific affinity for double-stranded DNA.

This article has been cited by other articles:

• Okishio, N, Adachi, Y, Yanagida, M (1996). Fission yeast Nda1 and Nda4, MCM homologs required for DNA replication, are constitutive nuclear proteins. J. Cell Sci. 109: 319-326 [Abstract]  
• Nabeshima, K, Kurooka, H, Takeuchi, M, Kinoshita, K, Nakaseko, Y, Yanagida, M (1995). p93dis1, which is required for sister chromatid separation, is a novel microtubule and spindle pole body-associating protein phosphorylated at the Cdc2 target sites.. Genes Dev. 9: 1572-1585 [Abstract]  
• Ohkura, H, Hagan, I M, Glover, D M (1995). The conserved Schizosaccharomyces pombe kinase plo1, required to form a bipolar spindle, the actin ring, and septum, can drive septum formation in G1 and G2 cells.. Genes Dev. 9: 1059-1073 [Abstract]  
• Yoshida, T., Toda, T., Yanagida, M. (1994). A calcineurin-like gene ppb1+ in fission yeast: mutant defects in cytokinesis, cell polarity, mating and spindle pole body positioning. J. Cell Sci. 107: 1725-1735 [Abstract]  
• Kato, G J, Lee, W M, Chen, L L, Dang, C V (1992). Max: functional domains and interaction with c-Myc.. Genes Dev. 6: 81-92 [Abstract]  
• Blackwood, E., Eisenman, R. (1991). Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science 251: 1211-1217 [Abstract]  
• Toda, T, Shimanuki, M, Yanagida, M (1991). Fission yeast genes that confer resistance to staurosporine encode an AP-1-like transcription factor and a protein kinase related to the mammalian ERK1/MAP2 and budding yeast FUS3 and KSS1 kinases.. Genes Dev. 5: 60-73 [Abstract]  
• Blackwell, T., Kretzner, L, Blackwood, E., Eisenman, R., Weintraub, H (1990). Sequence-specific DNA binding by the c-Myc protein. Science 250: 1149-1151 [Abstract]  
• Boldogh, I, AbuBakar, S, Albrecht, T (1990). Activation of proto-oncogenes: an immediate early event in human cytomegalovirus infection. Science 247: 561-564 [Abstract]  
• Miyatani, S, Shimamura, K, Hatta, M, Nagafuchi, A, Nose, A, Matsunaga, M, Hatta, K, Takeichi, M (1989). Neural cadherin: role in selective cell-cell adhesion. Science 245: 631-635 [Abstract]  
• DePinho, R A, Hatton, K S, Tesfaye, A, Yancopoulos, G D, Alt, F W (1987). The human myc gene family: structure and activity of L-myc and an L-myc pseudogene.. Genes Dev. 1: 1311-1326 [Abstract]  
• Studzinski, G., Brelvi, Z., Feldman, S., Watt, R. (1986). Participation of c-myc protein in DNA synthesis of human cells. Science 234: 467-470 [Abstract]  
• Rogers, S, Wells, R, Rechsteiner, M (1986). Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234: 364-368 [Abstract]  
• Kaczmarek, L, Hyland, J., Watt, R, Rosenberg, M, Baserga, R (1985). Microinjected c-myc as a competence factor. Science 228: 1313-1315 [Abstract]