MCB
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chen, Y.
Right arrow Articles by Pilz, R. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, Y.
Right arrow Articles by Pilz, R. B.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, June 2003, p. 4066-4082, Vol. 23, No. 12
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.12.4066-4082.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Synergism between Calcium and Cyclic GMP in Cyclic AMP Response Element-Dependent Transcriptional Regulation Requires Cooperation between CREB and C/EBP-ß

Yongchang Chen, Shunhui Zhuang, Stijn Cassenaer, Darren E. Casteel, Tanima Gudi, Gerry R. Boss, and Renate B. Pilz*

Department of Medicine and Cancer Center, University of California at San Diego, La Jolla, California 92093-0652

Received 24 September 2002/ Returned for modification 18 November 2002/ Accepted 18 March 2003

Calcium induces transcriptional activation of the fos promoter by activation of the cyclic AMP response element (CRE)-binding protein (CREB), and in some cells its effect is enhanced synergistically by cyclic GMP (cGMP) through an unknown mechanism. We observed calcium-cGMP synergism in neuronal and osteogenic cells which express type II cGMP-dependent protein kinase (G-kinase); the effect on the fos promoter was mediated by the CRE and proportional to G-kinase activity. Dominant negative transcription factors showed involvement of CREB- and C/EBP-related proteins but not of AP-1. Expression of C/EBP-ß but not C/EBP-{alpha} or -{delta} enhanced the effects of calcium and cGMP on a CRE-dependent reporter gene. The transactivation potential of full-length CREB fused to the DNA-binding domain of Gal4 was increased synergistically by calcium and cGMP, and overexpression of C/EBP-ß enhanced the effect, while a dominant negative C/EBP inhibited it. With a mammalian two-hybrid system, coimmunoprecipitation experiments, and in vitro binding studies, we demonstrated that C/EBP-ß and CREB interacted directly; this interaction involved the C terminus of C/EBP-ß but occurred independently of CREB's leucine zipper domain. CREB Ser133 phosphorylation was stimulated by calcium but not by cGMP; in cGMP-treated cells, 32PO4 incorporation into C/EBP-ß was decreased and C/EBP-ß/CRE complexes were increased, suggesting regulation of C/EBP-ß functions by G-kinase-dependent dephosphorylation. C/EBP-ß and CREB associated with the fos promoter in intact cells, and the amount of promoter-associated C/EBP-ß was increased by calcium and cGMP. We conclude that calcium and cGMP transcriptional synergism requires cooperation of CREB and C/EBP-ß, with calcium and cGMP modulating the phosphorylation states of CREB and C/EBP-ß, respectively.


* Corresponding author. Mailing address: Department of Medicine and Cancer Center, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0652. Phone: (858) 534-8805. Fax: (858) 534-1421. E-mail: rpilz{at}ucsd.edu.


Molecular and Cellular Biology, June 2003, p. 4066-4082, Vol. 23, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.12.4066-4082.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




This article has been cited by other articles:




Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. J. Virol. Eukaryot. Cell
Microbiol. Mol. Biol. Rev. Clin. Vaccine Immunol. All ASM Journals

Copyright © 2003 by the American Society for Microbiology. All rights reserved.