Phosphorylation of cyclic adenosine 3′,5′-monophosphate (cAMP) response element-binding protein isoforms by the cAMP-dependent protein kinase

Peiqing Sun, William E. Schoderbek, Richard Maurer

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The cAMP response element-binding protein (CREB) mediates transcriptional activation of genes in response to the cAMP signal transduction pathway. There are two different isoforms of CREB, which are generated by alternative RNA splicing. There is evidence that the two isoforms may have different biological activities. As the longer isoform (CREB341) contains a potential phosphorylation site that is not present in the shorter isoform (CREB327), we examined the possible differential phosphorylation of the two CREB isoforms. Recombinant CREB was prepared and used as substrate for phosphorylation by the cAMP-dependent protein kinase in vitro. Phosphopeptide mapping and mutagenesis studies demonstrated that CREB341 contains two sites, serine 133 and serine 98, that can be phosphorylated in vitro by the catalytic subunit of the cAMP-dependent protein kinase. In contrast, CREB327 contains only a single phosphorylation site at serine 119 (equivalent position to serine 133 in CREB341). A kinase titration experiment demonstrated that serine 98 of CREB341 was phosphorylated only at relatively high concentrations of the cAMP-dependent protein kinase. Transient transfection studies were used to test for any possible function of the phosphorylation of serine 98 of CREB341. These studies used GAL4-CREB fusion proteins. We found that mutation of serine 98 to alanine (which would block phosphorylation) has little or no effect on the ability of the CREB fusion protein to activate transcription. These findings suggest that differences in the biological activity of the two CREB isoforms are probably not mediated by differential phosphorylation by the cAMP-dependent protein ki-nase.

Original languageEnglish (US)
Pages (from-to)1858-1866
Number of pages9
JournalMolecular Endocrinology
Volume6
Issue number11
StatePublished - 1992
Externally publishedYes

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Cyclic AMP Response Element-Binding Protein
Response Elements
Adenosine
Protein Kinases
Serine
Carrier Proteins
Protein Isoforms
Phosphorylation
Phosphopeptides
Proteins
Alternative Splicing
Recombinant Proteins
Mutagenesis
Alanine
Transcriptional Activation
Transfection
Signal Transduction
Catalytic Domain
Phosphotransferases
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Phosphorylation of cyclic adenosine 3′,5′-monophosphate (cAMP) response element-binding protein isoforms by the cAMP-dependent protein kinase. / Sun, Peiqing; Schoderbek, William E.; Maurer, Richard.

In: Molecular Endocrinology, Vol. 6, No. 11, 1992, p. 1858-1866.

Research output: Contribution to journalArticle

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