Mitogen-Activated Protein (MAP) Kinase Phosphorylation of MAP Kinase Kinase: Determination of Phosphorylation Sites by Mass Spectrometry and Site-Directed Mutagenesis1

Mitogen-activated protein (MAP) kinase in human eggs has been investigated by using immunoblotting with both anti-Active MAPK and anti-ERK2 antibodies. The results showed that the main form of MAP kinase was p42ERK2. It was in a dephosphorylated form in oocytes at the germinal vesicle stage, but fully phosphorylated in unfertilised mature eggs. MAP kinase phosphorylation was significantly decreased when pronuclei were formed after intracytoplasmic sperm injection. Neither MAP kinase expression nor activity was detected in morphologically degenerated eggs. Although MAP kinase still existed in early embryos arrested at the 8-cell or morula stages, little, if any, activity could be detected. These data suggest that MAP kinase may play an important role in the cell cycle regulation of human eggs, as in other mammalian species.

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The Mitogen-activated Protein (MAP) Kinase p38 and Its Upstream Activator MAP Kinase Kinase 6 Are Involved in the Activation of Signal Transducer and Activator of Transcription by Hyperosmolarity

Journal of Biological Chemistry ◽

10.1074/jbc.274.42.30222 ◽

1999 ◽

Vol 274 (42) ◽

pp. 30222-30227 ◽

Cited By ~ 79

Author(s):

Johannes G. Bode ◽

Petros Gatsios ◽

Stephan Ludwig ◽

Ulf R. Rapp ◽

Dieter Häussinger ◽

...

Keyword(s):

Map Kinase ◽

Signal Transducer ◽

Protein Map ◽

Mitogen Activated Protein ◽

Map Kinase Kinase

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Inhibition by adenylyl cyclase-cAMP system of ET-1-induced HSP27 in osteoblasts

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.2001.281.6.e1260 ◽

2001 ◽

Vol 281 (6) ◽

pp. E1260-E1266 ◽

Cited By ~ 4

Author(s):

Daijiro Hatakeyama ◽

Osamu Kozawa ◽

Masayuki Niwa ◽

Hiroyuki Matsuno ◽

Kanefusa Kato ◽

...

Keyword(s):

Map Kinase ◽

Adenylyl Cyclase ◽

Mrna Level ◽

P38 Map Kinase ◽

Hsp 27 ◽

Kinase C ◽

Protein Map ◽

Mitogen Activated Protein ◽

Camp System ◽

Kinase Phosphorylation

We have previously reported that endothelin-1 (ET-1) stimulates heat shock protein (HSP) 27 induction in osteoblast-like MC3T3-E1 cells and that p38 mitogen-activated protein (MAP) kinase acts at a point downstream from protein kinase C (PKC) in HSP27 induction. In the present study, we investigated the effect of the adenylyl cyclase-cAMP system on ET-1-stimulated induction of HSP27 in MC3T3-E1 cells. Dibutyryl-cAMP (DBcAMP) dose dependently inhibited the HSP27 accumulation stimulated by ET-1. Forskolin and cholera toxin significantly suppressed the ET-1-stimulated accumulation of HSP27. However, dideoxyforskolin, a forskolin derivative that does not activate cAMP, failed to suppress the ET-1-induced HSP27 accumulation. Forskolin reduced the p38 MAP kinase phosphorylation induced by ET-1 or 12- O-tetradecanoylphorbol-13-acetate (TPA). PGE1, an extracellular agonist that activates cAMP production, reduced the ET-1-induced HSP27 accumulation. In addition, the phosphorylation of p38 MAP kinase induced by ET-1 or TPA was suppressed by PGE1. Forskolin, DBcAMP, and PGE1suppressed the ET-1-stimulated increase in the mRNA level for HSP27. These results indicate that the adenylyl cyclase-cAMP system has an inhibitory role in ET-1-stimulated HSP27 induction in osteoblasts and that the effect is exerted at the point between PKC and p38 MAP kinase in osteoblasts.

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cAMP inhibits mitogen-activated protein (MAP) kinase activation and resumption of meiosis, but exerts no effects after spontaneous germinal vesicle breakdown (GVBD) in mouse oocytes

Reproduction Fertility and Development ◽

10.1071/rd99038 ◽

1999 ◽

Vol 11 (2) ◽

pp. 81 ◽

Cited By ~ 23

Author(s):

Q. Y. Sun ◽

Q. Lu ◽

H. Breitbart ◽

D. Y. Chen

Keyword(s):

Map Kinase ◽

Map Kinases ◽

Specific Inhibitor ◽

Germinal Vesicle ◽

Germinal Vesicle Breakdown ◽

Kinase Activation ◽

Protein Map ◽

Mitogen Activated Protein ◽

Kinase Phosphorylation ◽

Pkc Activation

Various signaling molecules have been implicated in the oocyte G2/MII transition, including protein kinase C (PKC), cAMP and mitogen-activated protein (MAP) kinases. However, the cross-talk among these signaling pathways has not been elucidated. The present study demonstrates that both germinal vesicle break down (GVBD) and MAP kinase phosphorylation (activation) are inhibited when intraoocyte cAMP is increased by treating the GV-intact oocytes with dibutyryl cyclic AMP (dbcAMP), forskolin, or isobutylmethylxanthine (IBMX). Okadaic acid, a specific inhibitor of protein phosphatase-1 and -2A, completely overcame this effect. Calphostin C, a specific inhibitor of PKC, accelerated both GVBD and MAP kinase phosphorylation, and this effect was attenuated by increased intraoocyte cAMP, whereas PKC activation inhibited these events. Once GVBD occurred, the progression of oocyte maturation and MAP kinase phosphorylation were independent of cAMP. These results indicate that an increase in intraoocyte cAMP, in synergy with PKC activation, initiates a cascade of events resulting in inhibition of MAP kinase phosphorylation and GVBD in the mouse oocyte.

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A protein factor for ras p21-dependent activation of mitogen-activated protein (MAP) kinase through MAP kinase kinase.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.90.3.975 ◽

1993 ◽

Vol 90 (3) ◽

pp. 975-979 ◽

Cited By ~ 44

Author(s):

T. Itoh ◽

K. Kaibuchi ◽

T. Masuda ◽

T. Yamamoto ◽

Y. Matsuura ◽

...

Keyword(s):

Map Kinase ◽

Protein Factor ◽

Protein Map ◽

Mitogen Activated Protein ◽

Ras P21 ◽

Map Kinase Kinase

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Involvement of Ras and Raf in the Gi-coupled acetylcholine muscarinic m2 receptor activation of mitogen-activated protein (MAP) kinase kinase and MAP kinase.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)36498-1 ◽

1993 ◽

Vol 268 (26) ◽

pp. 19196-19199

Author(s):

S Winitz ◽

M Russell ◽

N.X. Qian ◽

A Gardner ◽

L Dwyer ◽

...

Keyword(s):

Map Kinase ◽

Receptor Activation ◽

Protein Map ◽

Mitogen Activated Protein ◽

Map Kinase Kinase

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Inhibition of p42 and p44 MAP kinase does not alter smooth muscle contraction in swine carotid artery

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1998.275.1.h131 ◽

1998 ◽

Vol 275 (1) ◽

pp. H131-H138 ◽

Cited By ~ 26

Author(s):

Isabelle Gorenne ◽

Xiaoling Su ◽

Robert S. Moreland

Keyword(s):

Smooth Muscle ◽

Muscle Contraction ◽

Map Kinase ◽

Map Kinases ◽

Kinase Activity ◽

Smooth Muscle Contraction ◽

Protein Map ◽

Mitogen Activated Protein ◽

Kinase Phosphorylation

Caldesmon inhibits myosin ATPase activity; phosphorylation of caldesmon reverses the inhibition. The caldesmon kinase is believed to be mitogen-activated protein (MAP) kinase. MAP kinases are activated during vascular stimulation, but a cause-and-effect relationship between kinase activity and contraction has not been established. We examined the role of MAP kinase in contraction using PD-098059, an inhibitor of MAP kinase kinase (MEK). MAP kinase activity was assessed using an anti-active MAP kinase antibody and direct measurement of MAP kinase catalyzed phosphorylation of myelin basic protein, MBP-(95—98). MAP kinase phosphorylation, stimulated by histamine (50 μM) or phorbol 12,13-dibutyrate (PDBu, 0.1 μM), was inhibited by PD-098059 (100 μM). PD-098059 did not alter the sensitivity or the maximal level of force in smooth muscle stimulated by histamine or PDBu, nor did PD-098059 affect contraction of β-escin-permeabilized tissue. Our data suggest that p44 and p42 MAP kinases are not involved in regulation of vascular smooth muscle contraction. These results do not, however, preclude a role for other isoforms of the MAP kinase family.

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