A highly active homeobox gene promoter regulated by Ets and Sp1 family members in normal granulosa cells and diverse tumor cell types

Manjeet K Rao, Sourindra Maiti, Honnavara N. Ananthaswamy, Miles F. Wilkinson

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

One mechanism by which normal cells become converted to tumor cells involves the aberrant transcriptional activation of genes that are normally silent. We characterize a promoter that normally exhibits highly tissue- and stage-specific expression but displays ubiquitous expression when cells become immortalized or malignant, regardless of their lineage or tissue origin. This promoter normally drives the expression of the Pem homeobox gene in specific cell types in ovary and placenta but is aberrantly expressed in lymphomas, neuroblastomas, retinoblastomas, carcinomas, and sarcomas. By deletion analysis we identified a region between nucleotides -80 and -104 that was absolutely critical for the expression from this distal Pem promoter (Pem Pd). Site-specific mutagenesis and transfection studies revealed that this region contains two consensus Ets sites and a single Sp1 site that were necessary for Pem Pd expression. Gel shift analysis showed that Ets and Sp1 family members bound to these sites. Transfection studies demonstrated that the Ets family members Elf1 and Gabp and the Sp1 family members Sp1 and Sp3 transactivated the Pem Pd. Surprisingly, we found that Sp3 was a more potent activator of the Pem Pd than was Sp1; this is unusual, because Sp3 is either a weak activator or a repressor of most other promoters. Activation by either Elf1 or Gabp required an intact Sp1 family member binding site, suggesting that Ets and Sp1 family members cooperate to activate Pem Pd transcription. Expression from the Pem Pd (either transiently transfected or endogenous) depended on the Ras pathway, which could explain both its Ets- and Sp1-dependent expression in normal cells and its aberrant expression in tumor cells, in which ras protooncogenes are frequently mutated. We suggest that the Pem Pd may be a useful model system to understand the molecular mechanism by which a tissue-specific promoter can be corrupted in tumor cells.

Original languageEnglish (US)
Pages (from-to)26036-26045
Number of pages10
JournalJournal of Biological Chemistry
Volume277
Issue number29
DOIs
StatePublished - Jul 19 2002
Externally publishedYes

Fingerprint

Granulosa Cell Tumor
Homeobox Genes
Tumors
Genes
Cells
Tissue
Chemical activation
Mutagenesis
Transcription
Nucleotides
Transfection
Gels
Binding Sites
Neoplasms
Retinoblastoma
Electrophoretic Mobility Shift Assay
Site-Directed Mutagenesis
Neuroblastoma
Sarcoma
Placenta

ASJC Scopus subject areas

  • Biochemistry

Cite this

A highly active homeobox gene promoter regulated by Ets and Sp1 family members in normal granulosa cells and diverse tumor cell types. / Rao, Manjeet K; Maiti, Sourindra; Ananthaswamy, Honnavara N.; Wilkinson, Miles F.

In: Journal of Biological Chemistry, Vol. 277, No. 29, 19.07.2002, p. 26036-26045.

Research output: Contribution to journalArticle

Rao, Manjeet K ; Maiti, Sourindra ; Ananthaswamy, Honnavara N. ; Wilkinson, Miles F. / A highly active homeobox gene promoter regulated by Ets and Sp1 family members in normal granulosa cells and diverse tumor cell types. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 29. pp. 26036-26045.
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