Aromatic hydrocarbon receptor interaction with the retinoblastoma protein potentiates repression of E2F-dependent transcription and cell cycle arrest

Alvaro Puga, Sonya J. Barnes, Timothy P. Dalton, Ching Yi Chang, Erik S. Knudsen, Michael A. Maier

Research output: Contribution to journalArticle

229 Citations (Scopus)

Abstract

Polyhalogenated aromatic hydrocarbons, of which 2,3,7,8-tetrachloro-p- dioxin (TCDD) is the prototype compound, elicit a variety of toxic, teratogenic, and carcinogenic responses in exposed animals and in humans. In cultured cells, TCDD shows marked effects on the regulation of cell cycle progression, including thymocyte apoptosis, induction of keratinocyte proliferation and terminal differentiation, and inhibition of estrogen- dependent proliferation in breast cancer cells. The presence of an LXCXE domain in the dioxin aromatic hydrocarbon receptor (AHR), suggested that the effects of TCDD on cell cycle regulation might be mediated by protein-protein interactions between AHR and the retinoblastoma protein (RB). Using the yeast two-hybrid system, AHR and RB were in fact shown to bind to each other. In vitro pull-down experiments with truncated AHR peptides indicated that at least two separate AHR domains form independent complexes with hypophosphorylated RB. Coimmunoprecipitation of whole cell lysates from human breast carcinoma MCF-7 cells, which express both proteins endogenously, revealed that AHR associates with RB in vivo only after receptor transformation and nuclear translocation. However, the AHR nuclear translocator and transcriptional heterodimerization partner, is not required for (nor is it a part of) the AHR-RB complexes detected in vitro. Ectopic expression of AHR and RB in human osteosarcoma SAOS-2 cells, which lack endogenous expression of both proteins, showed that AHR synergizes with RB to repress E2F-dependent transcription and to induce cell cycle arrest. Furthermore, AHR partly blocked T-antigen-mediated reversal of RB-dependent transcriptional repression. These results uncover a potential function for the AHR in cell cycle regulation and suggest that this function may be that of serving as an environmental sensor that signals cell cycle arrest when cells are exposed to certain environmental toxicants.

Original languageEnglish (US)
Pages (from-to)2943-2950
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number4
DOIs
StatePublished - Jan 28 2000

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Aromatic Hydrocarbons
Retinoblastoma Protein
Transcription
Cell Cycle Checkpoints
Cells
Cell Cycle
Cytoplasmic and Nuclear Receptors
Proteins
Breast Neoplasms
Two-Hybrid System Techniques
Dioxins
Peptide Receptors
Poisons
Viral Tumor Antigens
MCF-7 Cells
Osteosarcoma
Thymocytes
Hybrid systems
Keratinocytes
Yeast

ASJC Scopus subject areas

  • Biochemistry

Cite this

Aromatic hydrocarbon receptor interaction with the retinoblastoma protein potentiates repression of E2F-dependent transcription and cell cycle arrest. / Puga, Alvaro; Barnes, Sonya J.; Dalton, Timothy P.; Chang, Ching Yi; Knudsen, Erik S.; Maier, Michael A.

In: Journal of Biological Chemistry, Vol. 275, No. 4, 28.01.2000, p. 2943-2950.

Research output: Contribution to journalArticle

Puga, Alvaro ; Barnes, Sonya J. ; Dalton, Timothy P. ; Chang, Ching Yi ; Knudsen, Erik S. ; Maier, Michael A. / Aromatic hydrocarbon receptor interaction with the retinoblastoma protein potentiates repression of E2F-dependent transcription and cell cycle arrest. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 4. pp. 2943-2950.
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