Camp dependent protein kinase activity in IL2-stimulated T lymphocytes

S. M. Belkowski, Michael B. Prystowsky

Research output: Contribution to journalArticle

Abstract

Interieukin 2 stimulates a new program of gene expression which drives T lymphocytes through the cell cycle. Proliferating cell nuclear antigen (PCNA) functions as a requisite processivity factor for DNA porymerase. PCNA expression is regulated transcriptionally in part though tandem CRE sequences in the promoter and CRE binding proteins. IL2 stimulates CREB phosphorylation in T lymphocytes. The most commonly identified mechanism for CREB phosphorylation is through the cAMP dependent protein kinase, PKA. Therefore, we examined PKA activity in the cloned T lymphocyte L2 following IL2 stimulation. Using a radioactive assay method with the phosphate receptor LRRASLG as a substrate, we observed an increase during the first six hours of stimulation followed by a decrease at the time of entry into S phase. A second ELISA-based assay system employing the phosphate receptor RRRVTSAARRS as a substrate yielded results that showed both a small early and larger late rise in PKA levels throughout the 48 hour time period assayed. The ELISA-based assay conditions employed revealed that most of the PKA was in an inactive form at the time of harvest and that all enzyme activity was inhibited by Pl6-22 a PKA specific inhibitor. However, the radioactive assay method showed a much higher level of PKA in an active form which was also inhibited by the Pl6-22. The different results obtained with the two assay systems suggest that there are two cAMP-inducible enzymes phosphorylating the peptide substrates which account for the activities in IL2-stimulated T cell lysates.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

Fingerprint

T-cells
protein kinases
Protein Kinases
Interleukin-2
Assays
T-lymphocytes
T-Lymphocytes
Proliferating Cell Nuclear Antigen
assays
proliferating cell nuclear antigen
Phosphorylation
Enzyme-Linked Immunosorbent Assay
Phosphates
phosphorylation
Substrates
enzyme-linked immunosorbent assay
phosphates
Enzymes
Cyclic AMP-Dependent Protein Kinases
S Phase

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Camp dependent protein kinase activity in IL2-stimulated T lymphocytes. / Belkowski, S. M.; Prystowsky, Michael B.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

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