Evidence for clonal heterogeneity of the expression of six protein kinase C isoforms in murine B and T lymphocytes.
Protein kinase C (PKC), which plays a pivotal role in lymphocyte activation, represents a homologous family of at least nine proteins. Seven genes that encode PKC proteins have been identified. Since the regulatory properties and substrate specificities of the isoforms are not identical in vitro, it is possible that each isoform plays a unique role in cell activation. Toward an understanding of the role of PKC isoforms in lymphocyte activation we have studied the expression of mRNA encoding six of the isoforms (alpha, beta, gamma, delta, epsilon, and zeta) in T cell clones and B cell lines. PKC isoform phenotyping was done by MAPPing using isoform-specific primers and slot-blot analyses of mRNA were performed using specific probes. T cell clones and B cell lines were determined to express levels of the delta, epsilon, and zeta isoforms of PKC that were detectable by MAPPing. Plasmacytomas did not express PKC-beta message detectable by MAPPing. Slot blot analyses and Western blot analyses with peptide-specific antibody confirmed that B cell plasmacytomas did not express PKC-beta mRNA or protein. T cell clones and B cell lines were similar in that none expressed PKC-gamma. In cells that expressed PKC isoforms that were detectable by the MAPPing protocol, there was heterogeneity in the relative abundance of isoform mRNA (PKC-delta and -beta) and protein (PKC-beta and -epsilon). Such diversity of isoform expression could be responsible for the differential responsiveness of lymphocyte clones to activating stimuli.
Terajima
J
J
Department of Molecular Immunology, Syntex Research, Palo Alto, California 94304.
Tsutsumi
A
A
Freire-Moar
J
J
Cherwinski
H M
HM
Ransom
J T
JT
D000818:Animals; D001402:B-Lymphocytes / Q000201:enzymology*; D002460:Cell Line; D008223:Lymphoma / Q000201:enzymology; D009101:Multiple Myeloma / Q000201:enzymology; D011493:Protein Kinase C / Q000032:analysis* / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis*; D013601:T-Lymphocytes / Q000201:enzymology*
D012333:RNA, Messenger; D011493:Protein Kinase C