The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B

R. Frank, J. Zhang, H. Uchida, S. Meyers, S. W. Hiebert, Stephen D Nimer

Research output: Contribution to journalArticle

195 Citations (Scopus)

Abstract

The t(8;21) translocation, commonly found in acute myelogenous leukemia (AML), generates a fusion protein containing N-terminal AML1 and C-terminal ETO amino acids. The human AML1 gene encodes several related proteins that specifically bind to the sequence TGT/cGGT, located in the promoter regions of a variety of hematopoietic growth factor genes. To examine the abilities of the AML1B protein (which contains 479 amino acids), a shorter AML1A isoform (which contains amino acids 1-250), and the AML1/ ETO fusion protein (which contains AMLlA amino acids 1-177) to stimulate transcription from the GM-CSF promoter, we performed co-transfection experiments in T cells using a human GM-CSF promoter-CAT reporter gene plasmid and expression vectors that contain the cDNAs for one of the above proteins. Our data demonstrate that AML1B, but not AML1A or AML1/ETO transactivates the GM-CSF promoter, requiring the TGTGGT sequence contained between base pairs -68 and -53. Furthermore, we show that AML1/ETO, but not AML1A, inhibits the ability of AMLlB to stimulate CAT expression. Electrophoretic mobility shift assays demonstrated the specific binding of AML1 proteins to the GM-CSF promoter TGTGGT sequence, which does not require GM-CSF sequences immediately upstream of this binding site. Our data support a role for AML1B as a transcriptional activator and establish that the AML1/ETO fusion protein can act as a dominant negative protein on the human GM-CSF promoter. Although AML1/ETO does not stimulate the transcription of GM-CSF, it may function by inhibiting the normal activity of AML1B in AML cells with the t(8;21) translocation.

Original languageEnglish
Pages (from-to)2666-2674
Number of pages9
JournalOncogene
Volume11
Issue number12
StatePublished - Dec 21 1995
Externally publishedYes

Fingerprint

Granulocyte-Macrophage Colony-Stimulating Factor
Transcriptional Activation
Proteins
Amino Acids
Acute Myeloid Leukemia
Core Binding Factor Alpha 2 Subunit
Electrophoretic Mobility Shift Assay
Reporter Genes
Genetic Promoter Regions
Base Pairing
Genes
Transfection
Intercellular Signaling Peptides and Proteins
Carrier Proteins
Protein Isoforms
Plasmids
Complementary DNA
Binding Sites
T-Lymphocytes
Gene Expression

Keywords

  • AML1/ETO
  • AML1B
  • GM-CSF promoter
  • Transcriptional activation

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Frank, R., Zhang, J., Uchida, H., Meyers, S., Hiebert, S. W., & Nimer, S. D. (1995). The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B. Oncogene, 11(12), 2666-2674.

The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B. / Frank, R.; Zhang, J.; Uchida, H.; Meyers, S.; Hiebert, S. W.; Nimer, Stephen D.

In: Oncogene, Vol. 11, No. 12, 21.12.1995, p. 2666-2674.

Research output: Contribution to journalArticle

Frank, R, Zhang, J, Uchida, H, Meyers, S, Hiebert, SW & Nimer, SD 1995, 'The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B', Oncogene, vol. 11, no. 12, pp. 2666-2674.
Frank R, Zhang J, Uchida H, Meyers S, Hiebert SW, Nimer SD. The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B. Oncogene. 1995 Dec 21;11(12):2666-2674.
Frank, R. ; Zhang, J. ; Uchida, H. ; Meyers, S. ; Hiebert, S. W. ; Nimer, Stephen D. / The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B. In: Oncogene. 1995 ; Vol. 11, No. 12. pp. 2666-2674.
@article{5d47254d9f694dfdaadea3eaebd321ce,
title = "The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B",
abstract = "The t(8;21) translocation, commonly found in acute myelogenous leukemia (AML), generates a fusion protein containing N-terminal AML1 and C-terminal ETO amino acids. The human AML1 gene encodes several related proteins that specifically bind to the sequence TGT/cGGT, located in the promoter regions of a variety of hematopoietic growth factor genes. To examine the abilities of the AML1B protein (which contains 479 amino acids), a shorter AML1A isoform (which contains amino acids 1-250), and the AML1/ ETO fusion protein (which contains AMLlA amino acids 1-177) to stimulate transcription from the GM-CSF promoter, we performed co-transfection experiments in T cells using a human GM-CSF promoter-CAT reporter gene plasmid and expression vectors that contain the cDNAs for one of the above proteins. Our data demonstrate that AML1B, but not AML1A or AML1/ETO transactivates the GM-CSF promoter, requiring the TGTGGT sequence contained between base pairs -68 and -53. Furthermore, we show that AML1/ETO, but not AML1A, inhibits the ability of AMLlB to stimulate CAT expression. Electrophoretic mobility shift assays demonstrated the specific binding of AML1 proteins to the GM-CSF promoter TGTGGT sequence, which does not require GM-CSF sequences immediately upstream of this binding site. Our data support a role for AML1B as a transcriptional activator and establish that the AML1/ETO fusion protein can act as a dominant negative protein on the human GM-CSF promoter. Although AML1/ETO does not stimulate the transcription of GM-CSF, it may function by inhibiting the normal activity of AML1B in AML cells with the t(8;21) translocation.",
keywords = "AML1/ETO, AML1B, GM-CSF promoter, Transcriptional activation",
author = "R. Frank and J. Zhang and H. Uchida and S. Meyers and Hiebert, {S. W.} and Nimer, {Stephen D}",
year = "1995",
month = "12",
day = "21",
language = "English",
volume = "11",
pages = "2666--2674",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "12",

}

TY - JOUR

T1 - The AML1/ETO fusion protein blocks transactivation of the GM-CSF promoter by AML1B

AU - Frank, R.

AU - Zhang, J.

AU - Uchida, H.

AU - Meyers, S.

AU - Hiebert, S. W.

AU - Nimer, Stephen D

PY - 1995/12/21

Y1 - 1995/12/21

N2 - The t(8;21) translocation, commonly found in acute myelogenous leukemia (AML), generates a fusion protein containing N-terminal AML1 and C-terminal ETO amino acids. The human AML1 gene encodes several related proteins that specifically bind to the sequence TGT/cGGT, located in the promoter regions of a variety of hematopoietic growth factor genes. To examine the abilities of the AML1B protein (which contains 479 amino acids), a shorter AML1A isoform (which contains amino acids 1-250), and the AML1/ ETO fusion protein (which contains AMLlA amino acids 1-177) to stimulate transcription from the GM-CSF promoter, we performed co-transfection experiments in T cells using a human GM-CSF promoter-CAT reporter gene plasmid and expression vectors that contain the cDNAs for one of the above proteins. Our data demonstrate that AML1B, but not AML1A or AML1/ETO transactivates the GM-CSF promoter, requiring the TGTGGT sequence contained between base pairs -68 and -53. Furthermore, we show that AML1/ETO, but not AML1A, inhibits the ability of AMLlB to stimulate CAT expression. Electrophoretic mobility shift assays demonstrated the specific binding of AML1 proteins to the GM-CSF promoter TGTGGT sequence, which does not require GM-CSF sequences immediately upstream of this binding site. Our data support a role for AML1B as a transcriptional activator and establish that the AML1/ETO fusion protein can act as a dominant negative protein on the human GM-CSF promoter. Although AML1/ETO does not stimulate the transcription of GM-CSF, it may function by inhibiting the normal activity of AML1B in AML cells with the t(8;21) translocation.

AB - The t(8;21) translocation, commonly found in acute myelogenous leukemia (AML), generates a fusion protein containing N-terminal AML1 and C-terminal ETO amino acids. The human AML1 gene encodes several related proteins that specifically bind to the sequence TGT/cGGT, located in the promoter regions of a variety of hematopoietic growth factor genes. To examine the abilities of the AML1B protein (which contains 479 amino acids), a shorter AML1A isoform (which contains amino acids 1-250), and the AML1/ ETO fusion protein (which contains AMLlA amino acids 1-177) to stimulate transcription from the GM-CSF promoter, we performed co-transfection experiments in T cells using a human GM-CSF promoter-CAT reporter gene plasmid and expression vectors that contain the cDNAs for one of the above proteins. Our data demonstrate that AML1B, but not AML1A or AML1/ETO transactivates the GM-CSF promoter, requiring the TGTGGT sequence contained between base pairs -68 and -53. Furthermore, we show that AML1/ETO, but not AML1A, inhibits the ability of AMLlB to stimulate CAT expression. Electrophoretic mobility shift assays demonstrated the specific binding of AML1 proteins to the GM-CSF promoter TGTGGT sequence, which does not require GM-CSF sequences immediately upstream of this binding site. Our data support a role for AML1B as a transcriptional activator and establish that the AML1/ETO fusion protein can act as a dominant negative protein on the human GM-CSF promoter. Although AML1/ETO does not stimulate the transcription of GM-CSF, it may function by inhibiting the normal activity of AML1B in AML cells with the t(8;21) translocation.

KW - AML1/ETO

KW - AML1B

KW - GM-CSF promoter

KW - Transcriptional activation

UR - http://www.scopus.com/inward/record.url?scp=0029616633&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029616633&partnerID=8YFLogxK

M3 - Article

C2 - 8545124

AN - SCOPUS:0029616633

VL - 11

SP - 2666

EP - 2674

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 12

ER -