scholarly journals Functional Proteomic Analysis of Long-term Growth Factor Stimulation and Receptor Tyrosine Kinase Coactivation in Swiss 3T3 Fibroblasts

2012 ◽  
Vol 11 (12) ◽  
pp. 1690-1708 ◽  
Author(s):  
Kohji Nagano ◽  
Akunna Akpan ◽  
Gayathri Warnasuriya ◽  
Steven Corless ◽  
Nick Totty ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Proteomic Analysis ◽  
3T3 Fibroblasts ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

scholarly journals Possible involvement of protein kinase C and calcium ion in growth factor-induced expression of c-myc oncogene in Swiss 3T3 fibroblasts.

1986 ◽  
Vol 261 (3) ◽  
pp. 1187-1192
Author(s):  
K Kaibuchi ◽  
T Tsuda ◽  
A Kikuchi ◽  
T Tanimoto ◽  
T Yamashita ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Growth Factor ◽  
Protein Kinase ◽  
Calcium Ion ◽  
Induced Expression ◽  
Myc Oncogene ◽  
3T3 Fibroblasts ◽  
Kinase C ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

scholarly journals Inhibition of mitogen-induced DNA synthesis by bafilomycin A1 in Swiss 3T3 fibroblasts

1996 ◽  
Vol 313 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Andrew J. SAURIN ◽  
Jane HAMLETT ◽  
Michael J. CLAGUE ◽  
Stephen R. PENNINGTON
Keyword(s):  
Growth Factor ◽  
Dna Synthesis ◽  
Kinase Activity ◽  
Bafilomycin A1 ◽  
Tyrosine Kinase Activity ◽  
3T3 Fibroblasts ◽  
Mitogenic Signalling ◽  
Epidermal Growth ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

Quiescent cells (in G0) can be stimulated to enter the cell cycle and proceed to DNA synthesis in S-phase by a wide range of growth factors and mitogens. Activation of cell-surface growth factor receptors with intrinsic protein tyrosine kinase activity initiates autophosphorylation of the receptors and subsequent activation of signal transduction cascades. After activation the receptors undergo ligand-induced internalization to endosomes, which become acidified by the action of a vacuolar H+-ATPase (V-ATPase). The extent to which vesicular acidification plays a role in mitogenic signalling by receptors with intrinsic tyrosine kinase activity remains unknown. Here we have shown that bafilomycin A1, a specific inhibitor of V-ATPase, inhibits endosome acidification and mitogen-induced DNA synthesis in Swiss 3T3 fibroblasts. Addition of bafilomycin A1 at successively later times during G1 progressively decreased the inhibition of DNA synthesis such that no inhibition was observed when bafilomycin A1 was added at the onset of S-phase. Bafilomycin A1 also induced a dramatic but reversible change in the morphology of Swiss 3T3 cells. However, the rapid activation of c-fos mRNA accumulation by epidermal growth factor and insulin was unaffected by bafilomycin A1. Together, the results suggest that activation of the V-ATPase plays an important role in the mitogenic signalling pathways that occur during the G1 phase of the cell cycle but is not required for the initial epidermal growth factor and insulin-evoked signalling events that lead to c-fos mRNA expression.

scholarly journals Focal adhesion kinase (p125FAK) and paxillin are substrates for sphingomyelinase-induced tyrosine phosphorylation in Swiss 3T3 fibroblasts

1996 ◽  
Vol 315 (3) ◽  
pp. 1035-1040 ◽  
Author(s):  
Takehiko SASAKI ◽  
Kaoru HAZEKI ◽  
Osamu HAZEKI ◽  
Michio UI ◽  
Toshiaki KATADA
Keyword(s):  
Tyrosine Kinase ◽  
Focal Adhesion Kinase ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesion ◽  
Kinase Activity ◽  
Tyrosine Kinase Activity ◽  
3T3 Fibroblasts ◽  
Kinase Cascade ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

We examined the effect of sphingomyelinase on tyrosine phosphorylation of intracellular proteins in mouse Swiss 3T3 fibroblasts. Incubation of the cells with bacterial sphingomyelinase resulted in the elevation of tyrosine phosphorylation of multiple cellular proteins of 190, 130, 120, 97 and 70 kDa within minutes. The 120 and 70 kDa tyrosine-phosphorylated peptides were identified as p125 focal adhesion kinase (p125FAK) and paxillin respectively by the use of specific antibodies against the proteins. Tyrosine kinase activity associated with anti-p125FAK immunoprecipitate was stimulated by incubation of cells with sphingomyelinase. Cytochalasin D, which selectively disrupts the network of actin filaments, inhibited sphingomyelinase-induced tyrosine phosphorylation of p125FAK and elevation of tyrosine kinase activity in the anti-p125FAK immunoprecipitates. Sphingomyelinase-induced phosphorylation of p125FAK was not inhibited by wortmannin, an inhibitor of phosphatidylinositol 3-kinase. This was in sharp contrast with a wortmannin-sensitive phosphorylation of p125FAK observed in platelet-derived growth factor (PGDF)-stimulated cells. Thus hydrolysis of sphingomyelin is considered to regulate the tyrosine kinase cascade including p125FAK and paxillin by a mechanism distinct from PDGF.

scholarly journals Multiple sources of sn-1,2-diacylglycerol in platelet-derived-growth-factor-stimulated Swiss 3T3 fibroblasts. Evidence for activation of phosphoinositidase C and phosphatidylcholine-specific phospholipase D

1991 ◽  
Vol 279 (2) ◽  
pp. 559-565 ◽  
Author(s):  
R Plevin ◽  
S J Cook ◽  
S Palmer ◽  
M J O Wakelam
Keyword(s):  
Growth Factor ◽  
Phospholipase D ◽  
Lag Time ◽  
Water Soluble ◽  
Platelet Derived Growth Factor ◽  
Second Phase ◽  
Multiple Sources ◽  
3T3 Fibroblasts ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

Platelet-derived growth factor (PDGF) stimulated sn-1,2-diacylglycerol (DAG) mass formation in Swiss 3T3 fibroblasts with a lag time of some 30 s. The response was biphasic, with the second phase being sustained over time. PDGF also stimulated the formation of Ins(1,4,5)P3 with a similar lag time to the DAG response, suggesting that DAG is derived from PtdIns(4,5)P2 hydrolysis at this time point. PDGF-stimulated phosphatidylcholine (PtdCho) hydrolysis in Swiss 3T3 fibroblasts, as measured by the formation of water-soluble choline metabolites and phosphatidylbutanol (PtdBut) accumulation, was by a phospholipase D (PLD)-catalysed pathway which was kinetically downstream of initial PtdIns(4,5)P2 hydrolysis. Accumulation of PtdBut increased up to 15 min, suggesting that PLD activity is not rapidly densitized in response to PDGF. The kinetics of PtdCho hydrolysis closely paralleled the second phase of DAG formation, strongly suggesting that during prolonged stimulation periods PtdCho is a major source of DAG in these cells. However, since PtdIns(4,5)P2 breakdown was also prolonged, PDGF-stimulated DAG may be derived from both phospholipids. Down-regulation of protein kinase C (PKC), by pre-treatment with phorbol 12-myristate 13-acetate, abolished both [3H]choline and [3H]PtdBut formation, suggesting that PLD-catalysed PtdCho hydrolysis may be dependent on PKC activation, supporting its dependence on prior PtdIns(4,5)P2 hydrolysis.

Inhibitors of Calcineurin Block Expression of Cyclins A and E Induced by Fibroblast Growth Factor in Swiss 3T3 Fibroblasts

1998 ◽  
Vol 353 (2) ◽  
pp. 374-378 ◽  
Author(s):  
Masahiro Tomono ◽  
Kotaro Toyoshima ◽  
Motohiro Ito ◽  
Hisao Amano ◽  
Zoltan Kiss
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Fibroblast Growth ◽  
3T3 Fibroblasts ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

Bombesin treatment enhances vasopressin receptors in Swiss 3T3 cells

1993 ◽  
Vol 265 (6) ◽  
pp. C1658-C1662 ◽  
Author(s):  
M. M. Civan ◽  
J. Sinnett-Smith ◽  
M. Bouzyk ◽  
E. Rozengurt
Keyword(s):  
Wide Spectrum ◽  
Membrane Receptors ◽  
Scatchard Analysis ◽  
Biological Processes ◽  
3T3 Cells ◽  
3T3 Fibroblasts ◽  
Vasopressin Receptors ◽  
Swiss 3T3 ◽  
Swiss 3T3 Fibroblasts

Cells possess receptors for multiple different peptides that regulate a wide spectrum of biological processes. Although examples of homologous and heterologous downregulation have been reported, relatively little is known about the interaction between different peptides in modulating cellular activities. Here we demonstrate that pretreatment of Swiss 3T3 fibroblasts with 10 nM bombesin for 48 h enhanced the 45Ca2+ efflux acutely stimulated by vasopressin. The effect was not reciprocal, since preincubation with vasopressin did not affect the bombesin-stimulated Ca2+ efflux. Measurement of displaceable [3H] vasopressin binding demonstrated that bombesin pretreatment increases the hormonal binding by 3.8 +/- 0.2-fold (SE; n = 14) measured at 37 degrees C or at 4 degrees C. Scatchard analysis at 4 degrees C indicated that the increased binding reflects an increase in the number of vasopressin receptors without any significant effect on the apparent affinity of binding. Furthermore, addition of cycloheximide completely prevented the increase in [3H] vasopressin binding induced by bombesin. We conclude that long-term bombesin pretreatment induces heterologous enhancement of vasopressin responsiveness by increasing the number of membrane receptors.

Sign in / Sign up

Export Citation Format

Share Document