Protein Kinase A Regulatory Subunit Type IIβ Directly Interacts with and Suppresses CREB Transcriptional Activity in Activated T Cells

Elliott, Michael R.; Tolnay, Máté; Tsokos, George C.

doi:10.4049/jimmunol.171.7.3636

Cited by 28 publications

(29 citation statements)

References 32 publications

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“…Immunoprecipitation-Confluent 10-cm plates with HEK293 cells transfected with appropriate constructs were washed twice in ice-cold PBS and harvested in a cold lysis buffer containing 50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 1 mM dithiothreitol, and Complete protease inhibitor mixture (Roche Applied Science) (53). Lysates were clarified by centrifugation at 10,000 ϫ g for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein

Johannessen

Delghandi

Rykx

et al. 2007

Journal of Biological Chemistry

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Protein kinase D (PKD), a family of serine/threonine kinases, can be activated by a multitude of stimuli in a protein kinase C-dependent or -independent manner. PKD is involved in signal transduction pathways controlling cell proliferation, apoptosis, motility, and protein trafficking. Despite its versatile functions, few genuine in vivo substrates for PKD have been identified. In this study we demonstrate that the transcription factor cAMPresponse element-binding protein (CREB) is a direct substrate for PKD. PKD1 and CREB interact in cells, and activated PKD1 provokes CREB phosphorylation at Ser-133 both in vitro and in vivo. A constitutive active mutant of PKD1 stimulates GAL4-CREB-mediated transcription in a Ser-133-dependent manner, activates CRE-responsive promoters, and increases the expression of CREB target genes. PKD1 also enhances transcription mediated by two other members of the CREB family, ATF-1 and CREM. Our results describe a novel mechanism for PKD-induced signaling through activation of the transcription factor CREB and suggest that stimulus-induced phosphorylation of CREB, reported to be mediated by protein kinase C, may involve downstream activated PKD.The mammalian PKD 3 family of serine/threonine kinases includes the isoforms PKD1 (mouse PKD and human PKC), PKD2, and PKD3 (also named PKC). PKD was originally considered to be a member of the PKC family (1, 2) but is now classified in the calcium/calmodulin-dependent kinase group based on sequence similarities in the kinase domain (3). The PKDs share a similar architecture consisting of a C-terminal catalytic domain, an N-terminal regulatory domain that encompasses two cysteine-rich regions (C1a and C1b), and a pleckstrin homology (PH) domain (4 -7). A comparison of the amino acid sequences of PKD1-3 reveals that the highest homology lies in the catalytic domain, followed by C1a, C1b, and PH domain, suggesting that isoform-specific functions may be due to the regulatory part of the kinase (8).PKD1 can be activated by several mechanisms. The most studied mechanism involves a sequential activation of phospholipase C that results in the generation of the second messengers inositol 1,4,5-triphosphate and diacylglycerol (DAG) and subsequent activation of the classical (␣, ␤I, ␤II, and ␥) and novel (␦, ⑀, , and ) PKC isoforms. Binding of DAG to the C1b domain of PKD1 directs its translocation to the plasma membrane where activated novel PKC phosphorylates PKD1 at Ser-744/748 in the activation loop, causing activation of the enzyme. The activated enzyme can be imported via its C1b motif into the nucleus, where it transiently accumulates before being exported to the cytosol through a CRM1-dependent nuclear export pathway that requires the PH domain of PKD (4 -7). Mutations and deletions in the regulatory domain induce activation of PKD to various extents, and the entire regulatory domain has an inhibitory effect on the kinase activity (9). Indeed, PKD can also be activated through caspase-mediated cleavage of the regulatory domain (10). G␤␥ subunits ...

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Section: Methodsmentioning

confidence: 99%

Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein

Johannessen

Delghandi

Rykx

et al. 2007

Journal of Biological Chemistry

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“…In SLE T cells, the documented decreased PKA I activity (18) should be expected to result in decreased pCREB levels. Also, the RIIb subunit of PKA II, which is abnormally translocated from the cytoplasm to the nucleus (19), binds CREB and limits its binding to the IL2 promoter (20).…”

Section: Introductionmentioning

confidence: 99%

Protein phosphatase 2A is a negative regulator of IL-2 production in patients with systemic lupus erythematosus

Katsiari

Kyttaris²,

Juang³

et al. 2005

Journal of Clinical Investigation

143

145

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“…The finding that RII␤ levels are abnormally high in the nuclei of T cells of some SLE patients (18) raised the possibility that this subunit might contribute to the immunopathogenesis of SLE by modifying gene expression and, therefore, effector functions such as cytokine production. Indeed, T cells overexpressing RII␤ exhibit partitioning of the subunit between the cytosol and nucleus, down-regulation of CREB transcriptional activity, and diminished c-fos promoter activity and cFos protein production (22). These results provide evidence that RII␤ acts as a repressor of CREB-dependent gene transcription in activated T cells.…”

mentioning

confidence: 73%

Down-Regulation of IL-2 Production in T Lymphocytes by Phosphorylated Protein Kinase A-RIIβ

Elliott

Shanks

Khan

et al. 2004

The Journal of Immunology

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The β isoform of the type II regulatory subunit (RIIβ) of protein kinase A suppresses CREB transcriptional activity and c-Fos production in T cells following activation via the TCR. Because CREB is an integral nuclear transcription factor for IL-2 production by T cells, we tested the hypothesis that RIIβ down-regulates IL-2 expression and IL-2 production in T cells. Stable transfection of RIIβ in Jurkat T cells led to an ∼90% reduction in IL-2 mRNA and IL-2 protein following T cell activation. The inhibition of IL-2 production was associated with phosphorylation of the RIIβ subunit at serine 114 (pRIIβ) and localization of pRIIβ in intranuclear clusters. A serine 114 phosphorylation-defective mutant, RIIβS114A, did not form these intranuclear clusters as well as wild-type RIIβ, and did not inhibit IL-2 mRNA and protein synthesis, indicating that serine 114 phosphorylation is required for both nuclear localization and down-regulation of IL-2 production by RIIβ. In contrast to its effect on IL-2, RIIβ induced constitutive up-regulation of CD154 mRNA and cell surface expression. Thus, pRIIβ differentially regulates gene expression following T cell activation. Unexpectedly, we also found that stable overexpression of another protein kinase A regulatory subunit, RIα, had the opposite effect on IL-2 expression, causing a 3- to 4-fold increase in IL-2 production following stimulation. In summary, our data demonstrate a novel mechanism by which serine 114 phosphorylation and nuclear localization of RIIβ controls the regulation of gene expression in T cells.

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Protein Kinase A Regulatory Subunit Type IIβ Directly Interacts with and Suppresses CREB Transcriptional Activity in Activated T Cells

Cited by 28 publications

References 32 publications

Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein

Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein

Protein phosphatase 2A is a negative regulator of IL-2 production in patients with systemic lupus erythematosus

Down-Regulation of IL-2 Production in T Lymphocytes by Phosphorylated Protein Kinase A-RIIβ

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