Comparison of the mode of action of ras p21 with those of protein kinases A and C in the stimulation of gene expression in NIH/3T3 cells

FEBS Letters ◽  
1990 ◽  
Vol 269 (1) ◽  
pp. 148-152 ◽  
Author(s):  
Naohisa Oku ◽  
Kozo Kaibuchi ◽  
Yasuo Fukumoto ◽  
Yuichi Hori ◽  
Hiroyuki Fujioka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Kinases ◽  
Mode Of Action ◽  
3T3 Cells ◽  
Ras P21 ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

scholarly journals Differential early viral gene expression in two stages of human papillomavirus type 16 DNA-induced malignant transformation.

1987 ◽  
Vol 7 (6) ◽  
pp. 2165-2172 ◽  
Author(s):  
S Yasumoto ◽  
J Doniger ◽  
J A DiPaolo
Keyword(s):  
Gene Expression ◽  
Human Papillomavirus ◽  
Malignant Transformation ◽  
Cell Lines ◽  
Viral Gene Expression ◽  
Viral Gene ◽  
3T3 Cells ◽  
Hpv 16 ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Human papillomavirus (HPV) type 16 DNA induces progressive transformation in NIH 3T3 cells. Two types of cell lines, PM3T3G0 and PM3T3Fo, were isolated by G418 or focus selection, respectively, after transfection of cells by a recombinant HPV 16 DNA carrying the neo gene. These cell lines exhibited distinct phenotypes compared with controls. Saturation densities of PM3T3G0 and PM3T3Fo lines were two- to three- and five- to sevenfold greater than that of control NIH 3T3 cells, respectively. Neither cell type required high serum for growth, in contrast to NIH 3T3 cells. PM3T3G0 lines were premalignant, whereas PM3T3Fo lines manifested tumorigenicity within 2 weeks. Subpopulations of three PM3T3G0 lines underwent progressive transformation as reflected by focus formation. Analysis of HPV 16-specific mRNA species demonstrated that high levels of early and late gene expression were detected in premalignant PM3T3G0 lines, whereas relatively low quantities of selected gene messages were expressed in malignant transformants. Thus, high levels of viral gene expression are not crucial for malignant transformation.

Ectopic Expression of Protein Kinase CβII, -δ, and -ϵ, but Not -βI or -ζ, Provide for Insulin Stimulation of Glucose Uptake in NIH-3T3 Cells

1999 ◽  
Vol 372 (1) ◽  
pp. 69-79 ◽  
Author(s):  
Denise R. Cooper ◽  
James E. Watson ◽  
Niketa Patel ◽  
Philip Illingworth ◽  
Mildred Acevedo-Duncan ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Glucose Uptake ◽  
Ectopic Expression ◽  
3T3 Cells ◽  
Insulin Stimulation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

Stimulation of cell growth and proliferation in NIH-3T3 cells by onion and garlic oils

1986 ◽  
Vol 2 (3) ◽  
pp. 369-378 ◽  
Author(s):  
Judith T. Zelikoff ◽  
Norman M. Atkins ◽  
Sidney Belman
Keyword(s):  
Cell Growth ◽  
3T3 Cells ◽  
Cell Growth And Proliferation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

scholarly journals Regulation of Calreticulin Gene Expression by Calcium

1997 ◽  
Vol 138 (3) ◽  
pp. 547-557 ◽  
Author(s):  
Mathilde Waser ◽  
Nasrin Mesaeli ◽  
Charlotte Spencer ◽  
Marek Michalak
Keyword(s):  
Gene Expression ◽  
Genomic Dna ◽  
Chloramphenicol Acetyltransferase ◽  
Ionophore A23187 ◽  
Mrna Levels ◽  
3T3 Cells ◽  
Transcription Analysis ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

We have isolated and characterized a 12-kb mouse genomic DNA fragment containing the entire calreticulin gene and 2.14 kb of the promoter region. The mouse calreticulin gene consists of nine exons and eight introns, and it spans 4.2 kb of genomic DNA. A 1.8-kb fragment of the calreticulin promoter was subcloned into a reporter gene plasmid containing chloramphenicol acetyltransferase. This construct was then used in transient and stable transfection of NIH/ 3T3 cells. Treatment of transfected cells either with the Ca2+ ionophore A23187, or with the ER Ca2+-ATPase inhibitor thapsigargin, resulted in a five- to sevenfold increase of the expression of chloramphenicol acetyltransferase protein. Transactivation of the calreticulin promoter was also increased by fourfold in NIH/3T3 cells treated with bradykinin, a hormone that induces Ca2+ release from the intracellular Ca2+ stores. Analysis of the promoter deletion constructs revealed that A23187- and thapsigargin-responsive regions are confined to two regions (−115 to −260 and −685 to −1,763) in the calreticulin promoter that contain the CCAAT nucleotide sequences. Northern blot analysis of cells treated with A23187, or with thapsigargin, revealed a fivefold increase in calreticulin mRNA levels. Thapsigargin also induced a fourfold increase in calreticulun protein levels. Importantly, we show by nuclear run-on transcription analysis that calreticulin gene transcription is increased in NIH/3T3 cells treated with A23187 and thapsigargin in vivo. This increase in gene expression required over 4 h of continuous incubation with the drugs and was also sensitive to treatment with cycloheximide, suggesting that it is dependent on protein synthesis. Changes in the concentration of extracellular and cytoplasmic Ca2+ did not affect the increased expression of the calreticulin gene. These studies suggest that stress response to the depletion of intracellular Ca2+ stores induces expression of the calreticulin gene in vitro and in vivo.

Platelet-derived growth factor stimulation of GTPase-activating protein tyrosine phosphorylation in control and c-H-ras-expressing NIH 3T3 cells correlates with p21ras activation

1992 ◽  
Vol 12 (9) ◽  
pp. 3903-3909
Author(s):  
C J Molloy ◽  
T P Fleming ◽  
D P Bottaro ◽  
A Cuadrado ◽  
S A Aaronson
Keyword(s):  
Growth Factor ◽  
Tyrosine Phosphorylation ◽  
Platelet Derived Growth Factor ◽  
Gtpase Activating Protein ◽  
3T3 Cells ◽  
Pdgf Stimulation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Sustained Increase ◽  
Nih 3T3 Cells

Platelet-derived growth factor (PDGF) stimulation of NIH 3T3 cells leads to the rapid tyrosine phosphorylation of the GTPase-activating protein (GAP) and an associated 64- to 62-kDa tyrosine-phosphorylated protein (p64/62). To assess the functions of these proteins, we evaluated their phosphorylation state in normal NIH 3T3 cells as well as in cells transformed by oncogenically activated v-H-ras or overexpression of c-H-ras genes. No significant GAP tyrosine phosphorylation was observed in unstimulated cultures, while PDGF-BB induced rapid tyrosine phosphorylation of GAP in all cell lines analyzed. In NIH 3T3 cells, we found that PDGF stimulation led to the recovery of between 37 and 52% of GAP molecules by immunoprecipitation with monoclonal antiphosphotyrosine antibodies. Furthermore, PDGF exposure led to a rapid and sustained increase in the levels of p21ras bound to GTP, with kinetics similar to those observed for GAP tyrosine phosphorylation. The PDGF-induced increases in GTP-bound p21ras in NIH 3T3 cells were comparable to the steady-state level observed in serum-starved c-H-ras-overexpressing transformants, conditions in which these cells maintained high rates of DNA synthesis. These results imply that the level of p21ras activation following PDGF stimulation of NIH 3T3 cells is sufficient to support mitogenic stimulation. Addition of PDGF to c-H-ras-overexpressing cells also resulted in a rapid and sustained increase in GTP-bound p21ras. In these cells GAP, but not p64/62, showed increased tyrosine phosphorylation, with kinetics similar to those observed for increased GTP-bound p21ras. All of these findings support a role for GAP tyrosine phosphorylation in p21ras activation and mitogenic signaling.

scholarly journals Microinjection of ras p21 induces a rapid rise in intracellular pH.

1987 ◽  
Vol 7 (5) ◽  
pp. 1984-1988 ◽  
Author(s):  
N Hagag ◽  
J C Lacal ◽  
M Graber ◽  
S Aaronson ◽  
M V Viola
Keyword(s):  
Intracellular Ph ◽  
Rapid Rise ◽  
3T3 Cells ◽  
Ph Change ◽  
Ras P21 ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Quiescent mouse NIH 3T3 cells responded to microinjection of activated ras p21 with a rapid and sustained rise in intracellular pH (approximately 0.17 pH units). The p21-induced pH change was inhibited by amiloride treatment or growth of cells in medium low in sodium, suggesting a role for the Na+/H+ antiporter. Amiloride was found to suppress p21-induced mitosis, also.

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