Previous Article | Next Article 
Mol Cell Biol. 1985 March; 5(3): 448-456
Expression and characterization of the human c-myc DNA-binding protein.
R A Watt,
A R Shatzman and
M Rosenberg
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.
Mol Cell Biol. 1985 March; 5(3): 448-456
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]
-
Littlewood, T.D., Evan, G.I.
(1990). The Role of myc Oncogenes in Cell Growth and Differentiation. ADR
4: 69-79
[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]
-
Sullivan, N.F., Watt, R.A., Delannoy, M.R., Green, C.L., Spector, D.L.
(1986). Colocalization of the myc Oncogene Protein and Small Nuclear Ribonucleoprotein Particles. Cold Spring Harb Symp Quant Biol
51: 943-947
[Abstract]
-
Kaczmarek, L, Hyland, J., Watt, R, Rosenberg, M, Baserga, R
(1985). Microinjected c-myc as a competence factor. Science
228: 1313-1315
[Abstract]