Cyclin A-CDK phosphorylates Sp1 and enhances Sp1-mediated transcription

Patrick Fojas De Borja, N. Keith Collins, Ping Du, Jane Azizkhan-Clifford, Maria Mudryj

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

Cyclin A-mediated activation of cyclin-dependent kinases (CDKs) is essential for cell cycle transversal. Cyclin A activity is regulated on several levels and cyclin A elevation in a number of cancers suggests a role in tumorigenesis. In the present study, we used a modified DNA binding site selection and PCR amplification procedure to identify DNA binding proteins that are potential substrates of cyclin A-CDK. One of the sequences identified is the Sp1 transcription factor binding site. Co-immunoprecipitation experiments show that cyclin A and Sp1 can interact physically. In vitro and in vivo phosphorylation studies indicate that cyclin A-CDK complexes can phosphorylate Sp1. The phosphorylation site is located in the N-terminal region of the protein. Cells overexpressing cyclin A have elevated levels of Sp1 DNA binding activity, suggesting that cyclin A-CDK-mediated phosphorylation augments Sp1 DNA binding properties. In co-transfection studies, cyclin A expression stimulated transcription from an Sp1-regulated promoter. Mutation of the phosphorylation site abrogated cyclin A-CDK-dependent phosphorylation, augmentation of Sp1 transactivation function and DNA binding activity.

Original languageEnglish (US)
Pages (from-to)5737-5747
Number of pages11
JournalEMBO Journal
Volume20
Issue number20
DOIs
StatePublished - Oct 15 2001

Fingerprint

Cyclin A
Cyclin-Dependent Kinases
Transcription
Phosphorylation
DNA
Binding Sites
Sp1 Transcription Factor
Site selection
DNA-Binding Proteins
Immunoprecipitation
Transcriptional Activation
Transfection
Amplification
Cell Cycle
Carcinogenesis
Chemical activation
Cells

Keywords

  • Cyclin A
  • Cyclin-dependent kinase
  • Sp1
  • Transcription

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

De Borja, P. F., Collins, N. K., Du, P., Azizkhan-Clifford, J., & Mudryj, M. (2001). Cyclin A-CDK phosphorylates Sp1 and enhances Sp1-mediated transcription. EMBO Journal, 20(20), 5737-5747. https://doi.org/10.1093/emboj/20.20.5737

Cyclin A-CDK phosphorylates Sp1 and enhances Sp1-mediated transcription. / De Borja, Patrick Fojas; Collins, N. Keith; Du, Ping; Azizkhan-Clifford, Jane; Mudryj, Maria.

In: EMBO Journal, Vol. 20, No. 20, 15.10.2001, p. 5737-5747.

Research output: Contribution to journalArticle

De Borja, PF, Collins, NK, Du, P, Azizkhan-Clifford, J & Mudryj, M 2001, 'Cyclin A-CDK phosphorylates Sp1 and enhances Sp1-mediated transcription', EMBO Journal, vol. 20, no. 20, pp. 5737-5747. https://doi.org/10.1093/emboj/20.20.5737
De Borja, Patrick Fojas ; Collins, N. Keith ; Du, Ping ; Azizkhan-Clifford, Jane ; Mudryj, Maria. / Cyclin A-CDK phosphorylates Sp1 and enhances Sp1-mediated transcription. In: EMBO Journal. 2001 ; Vol. 20, No. 20. pp. 5737-5747.
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