The role of the transcription factor Sp1 in regulating the expression of the WAF1/CIP1 gene in U937 leukemic cells

Joseph R. Biggs, Jeffery E. Kudlow, Andrew Kraft

Research output: Contribution to journalArticle

198 Citations (Scopus)

Abstract

The Waf1/Cip1 protein induces cell cycle arrest through inhibition of the activity of cyclin-dependent kinases and proliferating cell nuclear antigen. Expression of the WAF1/CIP1 gene is induced in a p53-dependent manner in response to DNA damage but can also be induced in the absence of p53 by agents such as growth factors, phorbol esters, and okadaic acid. WAF1/CIP1 expression in U937 human leukemic cells is induced by both phorbol ester, a protein kinase C activator, and by okadaic acid, an inhibitor of phosphatases 1 and 2A. Both of these agents induce the differentiation of these leukemic cells toward macrophages. We demonstrate that phorbol esters and okadaic acid stimulate transcription from the WAF1/CIP1 promoter in U937 cells. This transcription is mediated by a region of the promoter between -154 and + 16, which contains two binding sites for the transcription factor Sp1. Deletion or mutation of these Sp1 sites reduces WAF1/CIP1 promoter response to phorbol ester and okadaic acid, while a reporter gene under the control of a promoter containing only multiple Sp1 binding sites and a TATA box is induced by phorbol ester and okadaic acid. The WAF1/CIP1 promoter is also highly induced by exogenous Sp1 in the Sp1-deficient Drosophila Schnieder SL 2 cell line. These results suggest that phorbol ester and okadaic acid activate transcription of the WAF1/CIP1 promoter through a complex of proteins that includes Sp1 and basal transcription factors.

Original languageEnglish (US)
Pages (from-to)901-906
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number2
DOIs
StatePublished - Jan 12 1996
Externally publishedYes

Fingerprint

Sp1 Transcription Factor
Okadaic Acid
U937 Cells
Phorbol Esters
Genes
Transcription
Binding Sites
Cells
Cell Cycle Proteins
TATA Box
Cyclin-Dependent Kinases
Macrophages
Sequence Deletion
Proliferating Cell Nuclear Antigen
Acid Phosphatase
Cell Cycle Checkpoints
Reporter Genes
Genetic Promoter Regions
Protein Kinase C
DNA Damage

ASJC Scopus subject areas

  • Biochemistry

Cite this

The role of the transcription factor Sp1 in regulating the expression of the WAF1/CIP1 gene in U937 leukemic cells. / Biggs, Joseph R.; Kudlow, Jeffery E.; Kraft, Andrew.

In: Journal of Biological Chemistry, Vol. 271, No. 2, 12.01.1996, p. 901-906.

Research output: Contribution to journalArticle

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