Linked Life Data

10064062

Source:http://linkedlifedata.com/resource/pubmed/id/10064062

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1999-3-16
pubmed:abstractText
Taking the antisense approach to inhibit the expression of specific protein kinase C (PKC) isoforms, we investigated the function of PKC alpha in T cell activation by transfecting Jurkat cells with an episomal vector (pREP3) containing a copy of the corresponding gene in the antisense orientation. Transfected Jurkat cells were selected with hygromycin and cloned by limiting dilution. Two (as1/as2) stably transfected antisense PKC alpha-pREP3 clones (as PKC alpha-pREP3) exhibited consistently reductions (76% and 85%, respectively) of PKC alpha levels when analyzed by immunoblotting and immunoprecipitation and also of PKC alpha mRNA (75%, as determined by Northern blotting) when compared to control clones (C1/C2) containing the pREP3 vector alone. The ability of the as-PKC alpha-pREP3 construct to specifically reduce PKC alpha levels in both clones was demonstrated by Western blots probed with antibodies against the PKC beta isozyme (the form structurally more similar to PKC alpha) and other representative isoenzymes expressed in Jurkat cells (PKC delta, epsilon, theta, and mu). Stimulation of transfected Jurkat clones with phorbol-12-myristate-13 alone or in the presence of ionomycin resulted in significant reduction of IL-2R alpha expression, TNF-alpha production, and the induction of transcriptional activity of a pIL-2/Luc construct in both as PKC alpha-reduced clones. The magnitude of these decrements paralleled the reductions of PKC alpha expression. The loss of the effects in clone as1 after a high number of passages correlated with the recovery of normal levels of PKC alpha protein, suggesting a link between these processes. Thus, the findings of this study demonstrate the essential role that PKC alpha plays in major events of the T lymphocyte activation process.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0014-2980
pubmed:author
pubmed:issnType
Print
pubmed:volume
29
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
466-76
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed:year
1999
pubmed:articleTitle
Inhibition of protein kinase C alpha expression by antisense RNA in transfected Jurkat cells.
pubmed:affiliation
Departamento de Bioquimica y Biologia Molecular, Facultad de Biologia, Universidad de Santiago de Compostela, España.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't