Identification and characterization of specific DNA-binding complexes containing members of the Myc/Max/Mad network of transcriptional regulators
In the past, eukaryotic cell-derived complexes of the Myc/Max/Mad network of transcriptional regulators have largely been refractory to DNA binding studies. We have developed electrophoretic mobility shift assay conditions to measure specific DNA binding of Myc/Max/Mad network complexes using a COS7 cell-based overexpression system. With the established protocol, we have measured on- and off-rates of c-Myc/Max, Max/Max, and Mad1/Max complexes and determined relative affinities. All three complexes appeared to bind with comparable affinity to a Myc E-box sequence. Furthermore, our data derived from competition experiments suggested that the Mad3/Max and Mad4/Max complexes also possess comparable DNA binding affinities. The conditions established for COS7 cell-overexpressed proteins were then used to identify c-Myc/Max, Max/Max, and Mnt/Max complexes in HL-60 cells. However, no Mad1/Max could be detected, despite the induction of Mad1 expression during differentiation. Whereas the DNA binding activity of c-Myc/Max complexes was down-regulated, Max/Max binding increased, and Mnt/Max binding remained unchanged. In addition, we have also tested for upstream stimulatory factor (USF) binding and observed that, in agreement with published data, USF comprises a major Myc E-box-binding factor that is more abundant than any of the Myc/Max/Mad network complexes. Similar to the Mnt/Max complex, the binding activity of USF remained constant during HL-60 differentiation. Our findings establish conditions for the analysis of DNA binding of Myc/Max/Mad complexes and indicate posttranslational regulation of the Max/Max complex.
| Medienart: |
Artikel |
|---|
| Erscheinungsjahr: |
1998 |
|---|---|
| Erschienen: |
1998 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:273 |
|---|---|
| Enthalten in: |
The Journal of biological chemistry - 273(1998), 12 vom: 20. März, Seite 6632-42 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Sommer, A [VerfasserIn] |
|---|
| Anmerkungen: |
Date Completed 16.04.1998 Date Revised 09.02.2021 published: Print Citation Status MEDLINE |
|---|
| Förderinstitution / Projekttitel: |
|
|---|
| PPN (Katalog-ID): |
NLM09448676X |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM09448676X | ||
| 003 | DE-627 | ||
| 005 | 20231222095758.0 | ||
| 007 | tu | ||
| 008 | 231222s1998 xx ||||| 00| ||eng c | ||
| 028 | 5 | 2 | |a pubmed24n0315.xml |
| 035 | |a (DE-627)NLM09448676X | ||
| 035 | |a (NLM)9506959 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Sommer, A |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Identification and characterization of specific DNA-binding complexes containing members of the Myc/Max/Mad network of transcriptional regulators |
| 264 | 1 | |c 1998 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 16.04.1998 | ||
| 500 | |a Date Revised 09.02.2021 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a In the past, eukaryotic cell-derived complexes of the Myc/Max/Mad network of transcriptional regulators have largely been refractory to DNA binding studies. We have developed electrophoretic mobility shift assay conditions to measure specific DNA binding of Myc/Max/Mad network complexes using a COS7 cell-based overexpression system. With the established protocol, we have measured on- and off-rates of c-Myc/Max, Max/Max, and Mad1/Max complexes and determined relative affinities. All three complexes appeared to bind with comparable affinity to a Myc E-box sequence. Furthermore, our data derived from competition experiments suggested that the Mad3/Max and Mad4/Max complexes also possess comparable DNA binding affinities. The conditions established for COS7 cell-overexpressed proteins were then used to identify c-Myc/Max, Max/Max, and Mnt/Max complexes in HL-60 cells. However, no Mad1/Max could be detected, despite the induction of Mad1 expression during differentiation. Whereas the DNA binding activity of c-Myc/Max complexes was down-regulated, Max/Max binding increased, and Mnt/Max binding remained unchanged. In addition, we have also tested for upstream stimulatory factor (USF) binding and observed that, in agreement with published data, USF comprises a major Myc E-box-binding factor that is more abundant than any of the Myc/Max/Mad network complexes. Similar to the Mnt/Max complex, the binding activity of USF remained constant during HL-60 differentiation. Our findings establish conditions for the analysis of DNA binding of Myc/Max/Mad complexes and indicate posttranslational regulation of the Max/Max complex | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Basic Helix-Loop-Helix Leucine Zipper Transcription Factors |2 NLM | |
| 650 | 7 | |a Basic-Leucine Zipper Transcription Factors |2 NLM | |
| 650 | 7 | |a DNA-Binding Proteins |2 NLM | |
| 650 | 7 | |a MAX protein, human |2 NLM | |
| 650 | 7 | |a MXD1 protein, human |2 NLM | |
| 650 | 7 | |a Myc associated factor X |2 NLM | |
| 650 | 7 | |a Proto-Oncogene Proteins c-myc |2 NLM | |
| 650 | 7 | |a Repressor Proteins |2 NLM | |
| 650 | 7 | |a Transcription Factors |2 NLM | |
| 650 | 7 | |a DNA |2 NLM | |
| 650 | 7 | |a 9007-49-2 |2 NLM | |
| 700 | 1 | |a Bousset, K |e verfasserin |4 aut | |
| 700 | 1 | |a Kremmer, E |e verfasserin |4 aut | |
| 700 | 1 | |a Austen, M |e verfasserin |4 aut | |
| 700 | 1 | |a Lüscher, B |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 273(1998), 12 vom: 20. März, Seite 6632-42 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:273 |g year:1998 |g number:12 |g day:20 |g month:03 |g pages:6632-42 |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 273 |j 1998 |e 12 |b 20 |c 03 |h 6632-42 | ||