Site‐selective 8‐C1‐cAMP which causes growth inhibition and differentiation increases DNA (CRE)‐binding activity in cancer cells

Mednieks, Maija I.; Yokozaki, Hiroshi; Merlo, Giorgio R.; Clair, Timothy; Ally, Shamsia; Tahara, Eiichi; Cho‐Chung, Yoon S.

doi:10.1016/0014-5793(89)81014-2

Cited by 13 publications

(1 citation statement)

References 25 publications

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“…8-N 3 -[ The double-stranded oligonucleotide containing a single octamer of CRE (TGACGTCA, underlined) 5Ј-AGAGATT-GCCTGACGTCAGAGAGCTAG-3Ј; Sp1, 5Ј-ATTCGATC-GGGGCGGGGCGAGC-3Ј; Oct-1, 5Ј-TGTCGAATGCAA-ATCACTAGAA-3Ј; and AP-1, 5Ј-CGCTTGATGAGTCAG-CCGGAA-3Ј were from Santa Cruz Biotechnology. The double-stranded oligonucleotide trioctamer of CRE 5ЈCCT-GACGTCATGACGTCATGACGTCA-3Ј was prepared as described (11). § These oligonucleotides were labeled with 32 P at the 5Ј end with T4 kinase (GIBCO͞BRL).…”

Section: Methodsmentioning

confidence: 99%

The RIIβ regulatory subunit of protein kinase A binds to cAMP response element: An alternative cAMP signaling pathway

Srivastava

Lee

Noguchi

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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ABSTRACTcAMP, through the activation of cAMPdependent protein kinase (PKA), is involved in transcriptional regulation. In eukaryotic cells, cAMP is not considered to alter the binding affinity of CREB͞ATF to cAMPresponsive element (CRE) but to induce serine phosphorylation and consequent increase in transcriptional activity. In contrast, in prokaryotic cells, cAMP enhances the DNA binding of the catabolite repressor protein to regulate the transcription of several operons. The structural similarity of the cAMP binding sites in catabolite repressor protein and regulatory subunit of PKA type II (RII) suggested the possibility of a similar role for RII in eukaryotic gene regulation. Herein we report that RII␤ subunit of PKA is a transcription factor capable of interacting physically and functionally with a CRE. In contrast to CREB͞ATF, the binding of RII␤ to a CRE was enhanced by cAMP, and in addition, RII␤ exhibited transcriptional activity as a Gal4-RII␤ fusion protein. These experiments identify RII␤ as a component of an alternative pathway for regulation of CRE-directed transcription in eukaryotic cells.cAMP-dependent protein kinase (PKA) is the major mediator of the cAMP signal transduction pathway in mammalian cells (1, 2). This enzyme consists of two catalytic (C) subunits and a regulatory (R) subunit dimer. Activation occurs when two cAMP molecules bind to each R subunit of PKA, resulting in the release of the C subunits.There are two types of PKA, type I (PKA-I) and type II (PKA-II), that share a common C subunit but contain different R subunits (RI and RII, respectively) (2). Through biochemical studies and gene cloning, four isoforms of the R subunits (RI␣, RI␤, RII␣, and RII␤) have been identified (3). Varying the ratios of two isoforms of PKA has been linked to cell growth and differentiation (4, 5). An enhanced expression of RI͞PKA-I correlates with active cell growth and cell transformation, whereas a decrease in RI͞PKA-I and an increase of RII͞PKA-II are related to growth inhibition and differentiation and͞or maturation (4, 5).Overexpression of the RII␤ subunit of PKA in several cancer cell lines results in a striking shift in PKA isozyme distribution, growth arrest, differentiation, and reverse transformation (4, 6, 7). The growth inhibition and reverse transformation correlated with nuclear translocation of RII␤, because the mutant RII␤ that failed to translocate into the nucleus was incapable of inducing reverse transformation (6).In this study, we examined whether RII␤ is a nuclear factor that can mediate cAMP responses in cAMP-responsive element (CRE)-containing genes in eukaryotic cells. Ki-rastransformed NIH 3T3 (DT) cells and DT cells infected with a retroviral vector containing the human RII␤ gene (8) (DTRII␤) or mutant RII␤-P (6) (DTRII␤-P) were used in the present study. The infectants were grown in the presence of 60 M ZnSO 4 for 48 hr before experiments to maximally induce the infected genes (6). On the basis of cell growth, ZnSO 4 treatment at 60 M for 5 days was not toxic to DT, DTRII...

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Section: Methodsmentioning

confidence: 99%

The RIIβ regulatory subunit of protein kinase A binds to cAMP response element: An alternative cAMP signaling pathway

Srivastava

Lee

Noguchi

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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Fast‐atom bombardment mass spectrometric analysis of cyclic nucleotide and nucleotide triphosphate analogues used in studies of cyclic nucleotide‐related enzymes

Langridge

Evans

Walton

et al. 1993

Rapid Comm Mass Spectrometry

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Two isomers of adenosine 3',5'-cyclic monophosphate, which show agonist and antagonist activity with cyclic AMP-dependent protein kinase, were found to yield essentially identical positive-ion fast-atom bombardment (FAB) mass spectra, but SI and S2 fragments of differing relative intensities in their collision-induced dissociations, studied using mass-analysed ion kinetic energy (CIDIMIKE) spectra. Halogen-substituted cyclic nucleotides, used in differentiating between protein kinase cyclic nucleotide binding sites, produced FAB mass spectra and CID/MIKE spectra with fragmentations generally analogous to those of the parent cyclic nucleotides; the bromo-derivatives showed a greater propensity for dehalogenation than the chloro-derivatives.The adenosine triphosphate analogues, adenylyl-(l), y-methylenel-diphosphate and adenylyl-imidodiphosphate, alternative substrates for adenylyl cyclase, showed similar fragmentations with the methylene and imido groups blocking cleavage between the l) and y phosphate groups. The fragmentations observed are discussed in the context of the use of these compounds in the assay of protein kinase and adenylyl cyclase activity by quantitative mass spectrometry.Fast-atom bombardment mass spectrometry (FAB-MS) in combination with collisionally induced dissociation and mass-analysed kinetic energy spectrometry (CID/MIKES) of quasi-molecular ions has provided an invaluable means of identifying putative cyclic nucleotides in tissue extracts'-3 and enzyme incubate^,^ and of determining the position and degree of substitution in cell-permeating synthetic derivatives.' The use of quantitative mass spectrometry has also been used to advantage in the study of enzymic reactions which function in the metabolism of cyclic nucleotide~~~' and in their mechanism of action as hormone mediators,8 and this approach has now been extended to the use of a continuous flow FAB which provides the basis for a continuous assay for these enzymes. In addition to enabling a continuous assay, not feasible by the conventional radiometric assays of these enzymes, the major advantage in application of quantitative mass spectrometric analysis to studies of enzyme activity is the ability to monitor several different reaction components simultaneously? and thus it is theoretically feasible to monitor, in the same incubation mixture, substrate(s), product(s), activator(s) and inhibitor(s). In view of the complexity of action of the cyclicnucleotide-related enzymes, the latter facility offers great potential; prior to systematic study of these enzymes by quantitative mass spectrometry, it is necessary first to establish the FAB-MS and CID/MIKES behaviour of compounds, additional to substrate and products previously examined, likely to be included in the enzyme incubates as putative effectors.Cyclic AMP-dependent protein kinase (1) is the enzyme that brings about the intracellular effect of cyclic AMP; it catalyses the phosphorylation of a range

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The Regulatory Subunit of cAMP-Dependent Protein Kinase as a Target for Cancer Diagnosis and Therapy

Cho‐Chung

Cereseto

Budillon

et al. 1993

Molecular Oncology and Clinical Applications

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Site‐selective 8‐C1‐cAMP which causes growth inhibition and differentiation increases DNA (CRE)‐binding activity in cancer cells

Cited by 13 publications

References 25 publications

The RIIβ regulatory subunit of protein kinase A binds to cAMP response element: An alternative cAMP signaling pathway

The RIIβ regulatory subunit of protein kinase A binds to cAMP response element: An alternative cAMP signaling pathway

Fast‐atom bombardment mass spectrometric analysis of cyclic nucleotide and nucleotide triphosphate analogues used in studies of cyclic nucleotide‐related enzymes

The Regulatory Subunit of cAMP-Dependent Protein Kinase as a Target for Cancer Diagnosis and Therapy

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