Activated or dominant inhibitory mutants of Rap1A decrease the oxidative burst of Epstein-Barr virus-transformed human B lymphocytes

Friedrich Ernst Maly, Lawrence Quilliam, Olivier Dorseuil, Channing J. Der, Gary M. Bokoch

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Abstract

Rap1A is a GTP-binding protein of the Ras superfamily that is highly abundant in phagocyte membranes. Although Rap1A copurifies with cytochrome b558, a component of the superoxide-generating NADPH oxidase complex of human phagocytes and B lymphocytes, the involvement of Rap1A in the regulation of the oxidative burst in these cells has not been clearly established. Therefore, we have stably transfected human Epstein-Barr virus- transformed B lymphocytes that possess an activable NADPH oxidase complex with cDNAs for mutants of Rap1A 'locked' in a GTP-bound (63E) and GDP-bound (17N) state. Both the 17N and 63E mutants of Rap1A inhibited phorbol ester- stimulated O2- production by 50 and 80%, respectively, while transfection with cDNA for wild-type Rap1A had no effect on the respiratory burst. No effects of the Rap1A mutants on cell viability, proliferation, expression of cell-surface markers, or phorbol 12-myristate 13-acetate- stimulated interleukin-8 generation were detected. These data demonstrate that Rap1A is a regulator of O2- formation in intact cells. Furthermore, the inhibitory effect of both GTP- as well as GDP-bound mutants indicates that Rap1A functions in a dynamic cycle as opposed to a unidirectional pathway, as is the case for the other NADPH oxidase regulatory GTP-binding protein, Rac.

Original languageEnglish (US)
Pages (from-to)18743-18746
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number29
StatePublished - Jul 22 1994
Externally publishedYes

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Respiratory Burst
Lymphocytes
NADPH Oxidase
Guanosine Triphosphate
Human Herpesvirus 4
Viruses
GTP-Binding Proteins
B-Lymphocytes
Complementary DNA
Phagocytes
Cell proliferation
Phorbol Esters
Interleukin-8
Acetates
Cells
Membranes
Transfection
Cell Survival
Cell Proliferation
phorbol-12-myristate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activated or dominant inhibitory mutants of Rap1A decrease the oxidative burst of Epstein-Barr virus-transformed human B lymphocytes. / Maly, Friedrich Ernst; Quilliam, Lawrence; Dorseuil, Olivier; Der, Channing J.; Bokoch, Gary M.

In: Journal of Biological Chemistry, Vol. 269, No. 29, 22.07.1994, p. 18743-18746.

Research output: Contribution to journalArticle

Maly, Friedrich Ernst ; Quilliam, Lawrence ; Dorseuil, Olivier ; Der, Channing J. ; Bokoch, Gary M. / Activated or dominant inhibitory mutants of Rap1A decrease the oxidative burst of Epstein-Barr virus-transformed human B lymphocytes. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 29. pp. 18743-18746.
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