Nitric oxide (NO) has gained increased attention as a diffusible universal messenger that plays a crucial role in the pathogenesis of inflammatory and autoimmune diseases. Recently, we reported that exogenous NO is able to activate the stress-activated protein kinase (SAPK) cascade in mesangial cells. Here, we demonstrate that exposure of glomerular mesangial cells to compounds releasing NO, including spermine-NO and (Z)-1-?N-methyl-N-[6-(N-methylammoniohexyl)amino]diazen?-1-ium+ ++-1,2-diolate (MAHMA-NO), results in an activation of the stress-activated p38-mitogen-activated protein kinase (p38-MAPK) cascade as measured by the phosphorylation of the activator of transcription factor-2 (ATF2) in an immunocomplex kinase assay. Activation of the p38-MAPK cascade by a short stimulation (10 min) with the NO donor MAHMA-NO causes a large increase in ATF2 phosphorylation that is several times greater than that observed after stimulation with interleukin-1beta, a well-known activator of the p38-MAPK pathway. Time course studies reveal that MAHMA-NO causes rapid and maximal activation of p38-MAPK after 10 min of stimulation and that activation declines to basal levels within 60 min. The longer-lived NO donor spermine-NO causes a comparable rapid activation of the p38-MAPK pathway; however, the increased activation state of p38-MAPK was maintained for several hours before control values were reattained after 24 h of stimulation. Furthermore, the NO donors also activated the classical extracellular signal-regulated kinase (ERK) p44-MAPK cascade as shown by phosphorylation of the specific substrate cytosolic phospholipase A2 in an immunocomplex kinase reaction. Both MAHMA-NO and spermine-NO cause a rapid activation of p44-MAPK after 10 min of stimulation. Interestingly, there is a second delayed peak of p44-MAPK activation after 4–24 h of stimulation with NO donors. These results suggest that there is a differential activation pattern for stress-activated and mitogen-activated protein kinases by NO and that the integration of these signals may lead to specific cell responses.

REFERENCES

Callsen
D.
,
Pfeilschifter
J.
,
Brune
B.
(
1998
).
Rapid and delayed p42/p44 MAPK activation by nitric oxide: the role of cGMP and tyrosine phosphatase inhibition
.
J. Immunol
161
,
4852
–.
Cano
E.
,
Mahadevan
L. C.
(
1995
).
Parallel signal processing among mammalian MAPKs
.
Trends Biochem. Sci
20
,
117
–.
Caselli
A.
,
Camici
G.
,
Manao
G.
,
Moneti
G.
,
Pazzagli
L.
,
Cappugi
G.
,
Ramponi
G.
(
1994
).
Nitric oxide causes inactivation of the low molecular weight phosphotyrosine protein phosphatase
.
J. Biol. Chem
269
,
24878
–.
Chuang
C.-F.
,
Ng
S.-Y.
(
1994
).
Functional divergence of the MAP kinase pathway. ERK1 and ERK2 activate specific transcription factors
.
FEBS Lett
346
,
229
–.
Cohen
J. J.
(
1993
).
Apoptosis
.
Immunol. Today
14
,
126
–.
Groom
L. A.
,
Sneddon
A. A.
,
Alessi
D. R.
,
Dowd
S.
,
Keyse
S. M.
(
1996
).
Differential regulation of the MAP, SAP and RK/p38 kinases by Pyst1, a novel cytosolic dual-specificity phosphatase
.
EMBO J
15
,
3621
–.
Huwiler
A.
,
Pfeilschifter
J.
(
1994
).
Extracellular ATP and UTP stimulate mitogen-activated protein kinase cascade and proliferation of rat renal mesangial cells
.
Br. J. Pharmac
113
,
1455
–.
Huwiler
A.
,
Pfeilschifter
J.
(
1994
).
Interleukin 1 stimulates de-novo synthesis of mitogen-activated protein kinase in glomerular mesangial cells
.
FEBS Lett
350
,
135
–.
Huwiler
A.
,
Stabel
S.
,
Fabbro
D.
,
Pfeilschifter
J.
(
1995
).
Platelet-derived growth factor and angiotensin II stimulate the mitogen-activated protein kinase cascade in rat renal mesangial cells
.
Biochem. J
305
,
777
–.
Keyse
S. M.
(
1995
).
An emerging family of dual specificity MAP kinase phosphatases
.
Biochim. Biophys. Acta
1265
,
152
–.
Kim
H.
,
Shim
J.
,
Han
P. L.
,
Choi
E. J.
(
1997
).
Nitric oxide modulates the c-Jun N-terminal kinase/stress-activated protein kinase activity through activating c-Jun N-terminal kinase kinase
.
Biochemistry
36
,
13677
–.
Knowles
R. G.
,
Moncada
S.
(
1994
).
Nitric oxide synthases in mammals
.
Biochem. J
298
,
249
–.
Lander
H. M.
,
Jacovina
A. T.
,
Davis
R. J.
,
Tauras
J. M.
(
1996
).
Differential activation of mitogen-activated protein kinases by nitric-oxide-related species
.
J. Biol. Chem
271
,
19705
–.
Lo
Y. Y. C.
,
Wong
J. M. S.
,
Cruz
T. F.
(
1996
).
Reactive oxygen species mediate cytokine activation of c-Jun NH2-terminal kinases
.
J. Biol. Chem
271
,
15703
–.
Meloche
S.
,
Seuwen
K.
,
Pages
G.
,
Pouyssegur
J.
(
1992
).
Biphasic and synergistic activation of p44mapk(ERK1) by growth factors: correlation between late phase activation and mitogenicity
.
Mol. Endocr
6
,
845
–.
Muhl
H.
,
Sandau
K.
,
Brune
B.
,
Briner
V. A.
,
Pfeilschifter
J.
(
1996
).
Nitric oxide donors induce apoptosis in glomerular mesangial cells, epithelial cells and endothelial cells
.
Eur. J. Pharmac
317
,
137
–.
Nathan
C.
(
1992
).
Nitric oxide as a secretory product of mammalian cells
.
FASEB J
6
,
3051
–.
Pfeilschifter
J.
(
1990
).
Extracellular ATP stimulates polyphosphoinositide hydrolysis and prostaglandin synthesis in rat renal mesangial cells. Involvement of a pertussis toxin-sensitive guanine nucleotide binding protein and feedback inhibition by protein kinase C
.
Cell Signal
2
,
129
–.
Pfeilschifter
J.
(
1990
).
Comparison of extracellular ATP and UTP signalling in rat renal mesangial cells. No indications for the involvement of separate purino-and pyrimidino-ceptors
.
Biochem. J
272
,
469
–.
Pfeilschifter
J.
(
1990
).
Angiotensin II B-type receptor mediates phosphoinositide hydrolysis in mesangial cells
.
Eur. J. Pharmac
184
,
201
–.
Pfeilschifter
J.
(
1995
).
Does nitric oxide, an inflammatory mediatorof glomerular mesangial cells, have a role in diabetic nephropathy?
.
Kidney Int
51
,
50
–.
Pfeilschifter
J.
,
Huwiler
A.
(
1996
).
Nitric oxide stimulates stress-activated protein kinases in glomerular endothelial and mesangial cells
.
FEBS Lett
396
,
67
–.
Pfeilschifter
J.
,
Pignat
W.
,
Vosbeck
K.
,
Märki
F.
(
1989
).
Interleukin 1 and tumor necrosis factor synergistically stimulate prostaglandin synthesis and phospholipase A2release from rat renal mesangial cells
.
Biochem. Biophys. Res. Commun
159
,
385
–.
Pfeilschifter
J.
,
Rob
P.
,
Mulsch
A.
,
Fandrey
J.
,
Vosbeck
K.
,
Busse
R.
(
1992
).
Interleukin 1 beta and tumour necrosis factor alpha induce a macrophage-type of nitric oxide synthase in rat renal mesangial cells
.
Eur. J. Biochem
203
,
251
–.
Pfeilschifter
J.
,
Schwarzenbach
H.
(
1990
).
Interleukin 1 and tumor necrosis factor stimulate cGMP formation in rat renal mesangial cells
.
FEBS Lett
273
,
185
–.
Stamler
J. S.
(
1994
).
Redox signaling: nitrosylation and related target interactions of nitric oxide
.
Cell
78
,
931
–.
Verheij
M.
,
Bose
R.
,
Lin
X. H.
,
Yao
B.
,
Jarvis
W. D.
,
Grant
S.
,
Birrer
M. J.
,
Szabo
E.
,
Zon
L. I.
,
Kyriakis
J. M.
,
Haimovitz-Friedman
A.
,
Fuks
Z.
,
Kolesnick
R. N.
(
1996
).
Requirement for ceramide-initiated SAPK/JNK signalling in stress-induced apoptosis
.
Nature
380
,
75
–.
Woodgett
J. R.
,
Kyriakis
J. M.
,
Avruch
J.
,
Zou
L. I.
,
Zanke
B.
,
Templeton
D. J.
(
1996
).
Reconstitution of novel signalling cascades responding to cellular stresses
.
Phil. Trans. R. Soc. Lond. B
351
,
135
–.
Xia
Z.
,
Dickens
M.
,
Raingeaud
J.
,
Davis
R. J.
,
Greenberg
M. E.
(
1995
).
Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis
.
Science
270
,
1326
–.
This content is only available via PDF.