PU.1 regulates the expression of the Vav proto-oncogene

Diane J. Denkinger, Taryn Q. Lambrecht, Allison M Cushman-Vokoun, Rodney S. Kawahara

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Vav is a guanine nucleotide exchange factor for the rho/rac GTPases that is upregulated in the embryo during the transition from primitive to definitive hematopoiesis. It is one of several genetic markers that correlates with the differentiation of the intraembryonic definitive hematopoietic stem cell. Subsequently, in the adult, vav is expressed predominantly in cells of the hematopoietic system. A heat-resistant protein complex that bound to a 23-bp segment, which is essential for vav promoter activity, was found to be present in myeloid cells but not T-cells. The complex was absent in non-hematopoietic cells which normally do not express vav. Using a saturation mutagenesis method, Mutex, a "footprint" of the protein binding site (AGAGGAAGT) was obtained that was consistent with the consensus binding site for PU.1. A specific antibody to PU.1 supershifted the complex and identified the presence of PU.1 within the complex. A GST fusion protein of the human PU.1 bound to the same consensus sequence as the heat-resistant complex from myeloid lineages. Specific mutation of the GGAA PU.1 core binding site silenced vav promoter activity and a dominant negative PU.1 inhibited the transactivation of PU.1 at the vav promoter as measured by the expression of the EGFP reporter gene. In addition, PCR analysis of immunoprecipitated chromatin using specific antibodies for PU.1 detected the co-immunoprecipitation of DNA containing the vav promoter. These results suggest that PU.1 is essential for transcriptional activity of the vav promoter in myeloid cells.

Original languageEnglish (US)
Pages (from-to)772-783
Number of pages12
JournalJournal of Cellular Biochemistry
Volume84
Issue number4
DOIs
StatePublished - Feb 11 2002

Fingerprint

Proto-Oncogenes
Binding Sites
Myeloid Cells
Hot Temperature
Rho Guanine Nucleotide Exchange Factors
Guanine Nucleotide Exchange Factors
Hematopoietic System
rho GTP-Binding Proteins
Mutagenesis
T-cells
Antibodies
GTP Phosphohydrolases
Hematopoiesis
Consensus Sequence
Hematopoietic Stem Cells
Stem cells
Genetic Markers
Reporter Genes
Immunoprecipitation
Protein Binding

Keywords

  • Development
  • Embryogenesis
  • Hematopoiesis
  • Stem cell
  • Transcription

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

PU.1 regulates the expression of the Vav proto-oncogene. / Denkinger, Diane J.; Lambrecht, Taryn Q.; Cushman-Vokoun, Allison M; Kawahara, Rodney S.

In: Journal of Cellular Biochemistry, Vol. 84, No. 4, 11.02.2002, p. 772-783.

Research output: Contribution to journalArticle

Denkinger, Diane J. ; Lambrecht, Taryn Q. ; Cushman-Vokoun, Allison M ; Kawahara, Rodney S. / PU.1 regulates the expression of the Vav proto-oncogene. In: Journal of Cellular Biochemistry. 2002 ; Vol. 84, No. 4. pp. 772-783.
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