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Journal Article | Research Support, Non-U.S. Gov't | Research Support, U.S. Gov't, P.H.S.

Protein kinase C-theta isoenzyme selective stimulation of the transcription factor complex AP-1 in T lymphocytes.

G Baier-Bitterlich, F Uberall, B Bauer, F Fresser, H Wachter, H Grunicke, G Utermann, A Altman, G Baier
G Baier-Bitterlich
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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F Uberall
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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B Bauer
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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F Fresser
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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H Wachter
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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H Grunicke
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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G Utermann
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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A Altman
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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G Baier
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
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DOI: 10.1128/MCB.16.4.1842
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ABSTRACT

T-lymphocyte stimulation requires activation of several protein kinases, including the major phorbol ester receptor protein kinase C (PKC), ultimately leading to induction of lymphokines, such as interleukin-2 (IL-2). The revelant PKC isoforms which are involved in the activation cascades of nuclear transcription factors involved in IL-2 production have not yet been clearly defined. We have examined the potential role of two representative PKC isoforms in the induction of the IL-2 gene, i.e., PKC-alpha and PKC-theta, the latter being expressed predominantly in hematopoietic cell lines, particularly T cells. Similar to that of PKC-alpha, PKC-theta overexpression in murine EL4 thymoma cells caused a significant increase in phorbol 12-myristate 13-acetate (PMA)-induced transcriptional activation of full-length IL-2-chloramphenicol acetyltransferase (CAT) and NF-AT-CAT but not of NF-IL2A-CAT or NF-kappaB promoter-CAT reporter gene constructs. Importantly, the critical AP-1 enhancer element was differentially modulated by these two distinct PKC isoenzymes, since only PKC-theta but not PKC-alpha overexpression resulted in an approximately 2.8-fold increase in AP-1-collagenase promoter CAT expression in comparison with the vector control. Deletion of the AP-1 enhancer site in the collagenase promoter rendered it unresponsive to PKC-theta. Expression of a constitutively active mutant PKC-theta A148E (but not PKC-alpha A25E) was sufficient to induce activation of AP-1 transcription factor complex in the absence of PMA stimulation. Conversely, a catalytically inactive PKC-theta K409R (but not PKC-alpha K368R) mutant abrogated endogenous PMA-mediated activation of AP-1 transcriptional complex. Dominant negative mutant Ha-RasS17N completely inhibited the PKC-O A148E-induced signal, PKC-O. Expression of a constitutively active mutant PKC-O A148E (but not PKC-alpha A25E) was sufficient to induce activation of AP-1 transcription factor complex in the absence of PMA stimulation. Conversely, a catalytically inactive PKC-O K409R (but not PKC-alpha K368R) mutant abrogated endogenous PMA-mediated activation of AP-1 transcriptional complex. Dominant negative mutant Ha-enRasS17N completely inhibited in the PKC-O A148E-induced signal, identifying PKC-theta as a specific constituent upstream of or parallel to Ras in the signaling cascade leading to AP transcriptional activation.

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Protein kinase C-theta isoenzyme selective stimulation of the transcription factor complex AP-1 in T lymphocytes.
G Baier-Bitterlich, F Uberall, B Bauer, F Fresser, H Wachter, H Grunicke, G Utermann, A Altman, G Baier
Molecular and Cellular Biology Apr 1996, 16 (4) 1842-1850; DOI: 10.1128/MCB.16.4.1842

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Protein kinase C-theta isoenzyme selective stimulation of the transcription factor complex AP-1 in T lymphocytes.
G Baier-Bitterlich, F Uberall, B Bauer, F Fresser, H Wachter, H Grunicke, G Utermann, A Altman, G Baier
Molecular and Cellular Biology Apr 1996, 16 (4) 1842-1850; DOI: 10.1128/MCB.16.4.1842
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