Kinetic analysis of the interaction of b/HLH/Z transcription factors Myc, Max, and Mad with cognate DNA.

Myc, Mad, and Max proteins belong to the basic helix-loop-helix leucine zipper family of transcription factors. They bind to a specific hexanucleotide element of DNA, the E-box (CACGTG). To be biologically active, Myc and Mad require dimerization with Max. For the route of complex assembly of these dimers, there are two proposed pathways. In the monomer pathway, two monomers bind DNA sequentially and assemble their dimerization interface while bound to DNA. In the dimer pathway, two monomers form a dimer first prior to association with DNA. The monomer pathway is kinetically favored. In this report, stopped-flow polarization was utilized to determine the rates and temperature dependence of all of the individual steps for both assembly pathways. Myc.Max dimerization had a rate constant approximately 5- and approximately 2-fold higher than those of Max.Max and Mad.Max dimerization, respectively. The protein dimerization rates as well as the dimer-DNA rates were found to be independent of concentration, suggesting conformational changes were rate-limiting. The Arrhenius activation energies for the dimerization of Myc, Mad, and Max with Max were 20.4 +/- 0.8, 29 +/- 0.6, and 40 +/- 0.2 kJ/mol, respectively. Further, rate constants for Max.Max homodimer DNA binding are significantly higher than for Myc.Max and Mad.Max heterodimers binding to DNA. Monomer-DNA binding showed a faster rate than dimer-DNA binding. These studies show the rate-limiting step for the dimer pathway is the formation of protein dimers, and this reaction is slower than formation of protein dimers on the DNA interface, kinetically favoring the monomer pathway.

[1]  R. Eisenman,et al.  Myc's broad reach. , 2008, Genes & development.

[2]  R. Cencic,et al.  c-Myc and eIF4F are components of a feedforward loop that links transcription and translation. , 2008, Cancer research.

[3]  J. Boult,et al.  Oesophageal adenocarcinoma is associated with a deregulation in the MYC/MAX/MAD network , 2008, British Journal of Cancer.

[4]  J. Gearhart,et al.  Pluripotency redux--advances in stem-cell research. , 2007, The New England journal of medicine.

[5]  I. Jelesarov,et al.  Thermodynamics of b-HLH-LZ protein binding to DNA: the energetic importance of protein-DNA contacts in site-specific E-box recognition by the complete gene product of the Max p21 transcription factor. , 2007, Biochemistry.

[6]  Pierre Lavigne,et al.  The mechanism of discrimination between cognate and non-specific DNA by dimeric b/HLH/LZ transcription factors. , 2007, Journal of molecular biology.

[7]  D. Goss,et al.  Interaction of Genome-linked Protein (VPg) of Turnip Mosaic Virus with Wheat Germ Translation Initiation Factors eIFiso4E and eIFiso4F* , 2006, Journal of Biological Chemistry.

[8]  S. Burley,et al.  Structural aspects of interactions within the Myc/Max/Mad network. , 2006, Current topics in microbiology and immunology.

[9]  D. Goss,et al.  Thermodynamics of protein-protein interactions of cMyc, Max, and Mad: effect of polyions on protein dimerization. , 2006, Biochemistry.

[10]  D. Goss,et al.  Assembly of b/HLH/z proteins c-Myc, Max, and Mad1 with cognate DNA: importance of protein-protein and protein-DNA interactions. , 2005, Biochemistry.

[11]  D. Goss,et al.  Translation initiation factor (eIF) 4B affects the rates of binding of the mRNA m7G cap analogue to wheat germ eIFiso4F and eIFiso4F.PABP. , 2005, Biochemistry.

[12]  B. Hogan,et al.  Nmyc plays an essential role during lung development as a dosage-sensitive regulator of progenitor cell proliferation and differentiation , 2005, Development.

[13]  J. Marko,et al.  How do site-specific DNA-binding proteins find their targets? , 2004, Nucleic acids research.

[14]  Chul-hak Yang,et al.  Determination of binding constant of transcription factor myc-max/max-max and E-box DNA: the effect of inhibitors on the binding. , 2004, Biochimica et biophysica acta.

[15]  Stephen K. Burley,et al.  X-Ray Structures of Myc-Max and Mad-Max Recognizing DNA Molecular Bases of Regulation by Proto-Oncogenic Transcription Factors , 2003, Cell.

[16]  R. Eisenman,et al.  Direct activation of RNA polymerase III transcription by c-Myc , 2003, Nature.

[17]  P. Farnham,et al.  c-Myc Mediates Activation of the cad Promoter via a Post-RNA Polymerase II Recruitment Mechanism* , 2001, The Journal of Biological Chemistry.

[18]  A. Schepartz,et al.  Effects of nucleic acids and polyanions on dimer formation and DNA binding by bZIP and bHLHZip transcription factors. , 2001, Bioorganic & medicinal chemistry.

[19]  M. Eilers,et al.  Regulation of cyclin D2 gene expression by the Myc/Max/Mad network: Myc-dependent TRRAP recruitment and histone acetylation at the cyclin D2 promoter. , 2001, Genes & development.

[20]  Jennifer J. Kohler,et al.  Kinetic studies of Fos.Jun.DNA complex formation: DNA binding prior to dimerization. , 2001, Biochemistry.

[21]  R. Eisenman,et al.  The Myc/Max/Mad network and the transcriptional control of cell behavior. , 2000, Annual review of cell and developmental biology.

[22]  A. Schepartz,et al.  DNA specificity enhanced by sequential binding of protein monomers. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Michael R. Green,et al.  Enhancement of TBP binding by activators and general transcription factors , 1999, Nature.

[24]  Robert T. Sauer,et al.  Acceleration of the refolding of Arc repressor by nucleic acids and other polyanions , 1999, Nature Structural Biology.

[25]  H. Dyson,et al.  DNA-induced conformational changes are the basis for cooperative dimerization by the DNA binding domain of the retinoid X receptor. , 1998, Journal of molecular biology.

[26]  T. Ellenberger,et al.  DNA-mediated Folding and Assembly of MyoD-E47 Heterodimers* , 1998, The Journal of Biological Chemistry.

[27]  T. Lee,et al.  Myc represses transcription of the growth arrest gene gas1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[28]  A. Schepartz,et al.  Certain bZIP peptides bind DMA sequentially as monomers and dimerize on the DMA , 1997, Nature Structural Biology.

[29]  M. Henriksson,et al.  Proteins of the Myc network: essential regulators of cell growth and differentiation. , 1996, Advances in cancer research.

[30]  N. Copeland,et al.  Mad3 and Mad4: novel Max-interacting transcriptional repressors that suppress c-myc dependent transformation and are expressed during neural and epidermal differentiation. , 1996, The EMBO journal.

[31]  K. Browning,et al.  Interaction of Wheat Germ Protein Synthesis Initiation Factor eIF-(iso)4F and Its Subunits p28 and p86 with m7GTP and mRNA Analogues (*) , 1995, The Journal of Biological Chemistry.

[32]  Steven L. Cohen,et al.  Probing the solution structure of the DNA‐binding protein Max by a combination of proteolysis and mass spectrometry , 1995, Protein science : a publication of the Protein Society.

[33]  Linheng Li,et al.  Regulation of cell proliferation and differentiation by Myc , 1995, Journal of Cell Science.

[34]  J. Darnell,et al.  Interferon activation of the transcription factor Stat91 involves dimerization through SH2-phosphotyrosyl peptide interactions , 1994, Cell.

[35]  Michael R. Green,et al.  Eukaryotic activators function during multiple steps of preinitiation complex assembly , 1993, Nature.

[36]  G. Evan,et al.  The c‐Myc protein induces cell cycle progression and apoptosis through dimerization with Max. , 1993, The EMBO journal.

[37]  B. Svensson,et al.  Reaction mechanisms of Trp120-->Phe and wild-type glucoamylases from Aspergillus niger. Interactions with maltooligodextrins and acarbose. , 1993, Biochemistry.

[38]  Stephen K. Burley,et al.  Recognition by Max of its cognate DNA through a dimeric b/HLH/Z domain , 1993, Nature.

[39]  R. Eisenman,et al.  Mad: A heterodimeric partner for Max that antagonizes Myc transcriptional activity , 1993, Cell.

[40]  R. Brent,et al.  Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. , 1993, Cell.

[41]  J. W. Little,et al.  Dimerization of a specific DNA-binding protein on the DNA. , 1992, Science.

[42]  G. Prendergast,et al.  Association of Myn, the murine homolog of Max, with c-Myc stimulates methylation-sensitive DNA binding and ras cotransformation , 1991, Cell.

[43]  D. Garland.  Kinetics and mechanism of colchicine binding to tubulin: evidence for ligand-induced conformational change. , 1978, Biochemistry.

[44]  P. Vogt,et al.  The RNA of avian acute leukemia virus MC29. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[46]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.