Loss of the amino-terminal helix-loop-helix domain of the vav proto-oncogene activates its transforming potential

1991 ◽  
Vol 11 (4) ◽  
pp. 1912-1920
Author(s):  
S Katzav ◽  
J L Cleveland ◽  
H E Heslop ◽  
D Pulido
Keyword(s):  
Leucine Zipper ◽  
Ectopic Expression ◽  
Cdna Clones ◽  
3T3 Cells ◽  
Amino Terminal ◽  
Helix Loop Helix ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

vav, a novel human oncogene, was originally generated in vitro by replacement of its normal 5' coding sequences with sequences from pSV2neo DNA, cotransfected as a selectable marker (S. Katzav, D. Martin-Zanca, and M. Barbacid, EMBO J. 8:2283-2290, 1989). The vav proto-oncogene is normally expressed in cells of hematopoietic origin. To determine whether the 5' rearrangement of vav or its ectopic expression in NIH 3T3 cells contributes to its transforming potential, we isolated murine and human proto-vav cDNA clones as well as human genomic clones corresponding to the 5' end of the gene. Normal proto-vav was poorly transforming in NIH 3T3 cells, whereas truncation of its 5' end greatly enhanced its transforming activity. The relative failure of full-length proto-vav cDNA clones to transform NIH 3T3 cells indicates that the transforming activity of vav is not simply due to ectopic expression. Analysis of the predicted amino terminus of the vav proto-oncogene shows that it contains a helix-loop-helix domain and a leucine zipper motif similar to that of myc family proteins, though it lacks a basic region that is usually found adjacent to helix-loop-helix domains. Loss of the helix-loop-helix domain of proto-vav, either by truncation or by rearrangement with pSV2neo sequences, activates its oncogenic potential.

scholarly journals Loss of the amino-terminal helix-loop-helix domain of the vav proto-oncogene activates its transforming potential.

1991 ◽  
Vol 11 (4) ◽  
pp. 1912-1920 ◽  
Author(s):  
S Katzav ◽  
J L Cleveland ◽  
H E Heslop ◽  
D Pulido
Keyword(s):  
Leucine Zipper ◽  
Ectopic Expression ◽  
Cdna Clones ◽  
3T3 Cells ◽  
Amino Terminal ◽  
Helix Loop Helix ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

vav, a novel human oncogene, was originally generated in vitro by replacement of its normal 5' coding sequences with sequences from pSV2neo DNA, cotransfected as a selectable marker (S. Katzav, D. Martin-Zanca, and M. Barbacid, EMBO J. 8:2283-2290, 1989). The vav proto-oncogene is normally expressed in cells of hematopoietic origin. To determine whether the 5' rearrangement of vav or its ectopic expression in NIH 3T3 cells contributes to its transforming potential, we isolated murine and human proto-vav cDNA clones as well as human genomic clones corresponding to the 5' end of the gene. Normal proto-vav was poorly transforming in NIH 3T3 cells, whereas truncation of its 5' end greatly enhanced its transforming activity. The relative failure of full-length proto-vav cDNA clones to transform NIH 3T3 cells indicates that the transforming activity of vav is not simply due to ectopic expression. Analysis of the predicted amino terminus of the vav proto-oncogene shows that it contains a helix-loop-helix domain and a leucine zipper motif similar to that of myc family proteins, though it lacks a basic region that is usually found adjacent to helix-loop-helix domains. Loss of the helix-loop-helix domain of proto-vav, either by truncation or by rearrangement with pSV2neo sequences, activates its oncogenic potential.

scholarly journals B-raf, a new member of the raf family, is activated by DNA rearrangement.

1988 ◽  
Vol 8 (6) ◽  
pp. 2651-2654 ◽  
Author(s):  
S Ikawa ◽  
M Fukui ◽  
Y Ueyama ◽  
N Tamaoki ◽  
T Yamamoto ◽  
...  
Keyword(s):  
Ewing Sarcoma ◽  
Dna Rearrangement ◽  
Terminal Portion ◽  
3T3 Cells ◽  
Complementary Dna ◽  
Amino Terminal ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Transforming Gene ◽  
Nih 3T3 Cells

Complementary DNA clones of a putative transforming gene were isolated from NIH 3T3 cells transformed with human Ewing sarcoma DNA. The gene was termed B-raf because it is related to but distinct from c-raf and A-raf. It appears that substitution in the amino-terminal portion of the normal B-raf protein confers transforming activity to the gene.

scholarly journals mos gene transforming efficiencies correlate with oocyte maturation and cytostatic factor activities.

1991 ◽  
Vol 11 (2) ◽  
pp. 604-610 ◽  
Author(s):  
N Yew ◽  
M Oskarsson ◽  
I Daar ◽  
D G Blair ◽  
G F Vande Woude
Keyword(s):  
Oocyte Maturation ◽  
Xenopus Oocytes ◽  
Kinase Activity ◽  
Vertebrate Species ◽  
3T3 Cells ◽  
Transforming Activity ◽  
Cytostatic Factor ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

The mos proto-oncogenes from different vertebrate species transform mouse NIH 3T3 cells with markedly different efficiencies. v-mos, mouse (c-mosmu), and chicken (c-mosch) mos transform NIH 3T3 cells 10- to 100-fold more efficiently than do human (c-moshu) and Xenopus (c-mosxc) mos. The mos genes with the highest transforming activity efficiently induce maturation in Xenopus oocytes and mimic cytostatic factor (CSF) by causing mitotic cleavage arrest in embryos. Chimeric v-mos/c-moshu proteins that had high transforming efficiencies in NIH 3T3 cells were also effective in the induction of oocyte maturation and CSF cleavage arrest. We measured the in vitro autophosphorylation activities of the different mos proteins and found that the levels of kinase activity of v-mos, c-mosmu, and c-mosch were much higher than that of c-mosxc. These data indicate that mos gene transforming efficiency and the ability to induce oocyte maturation or mimic CSF activity are correlated with in vitro autophosphorylation activity and suggest that the mos protein plays a similar role in transformed cells and normal oocytes.

scholarly journals Regulation of the cfos serum response element by C/EBPbeta.

1997 ◽  
Vol 17 (3) ◽  
pp. 1744-1755 ◽  
Author(s):  
L Sealy ◽  
D Malone ◽  
M Pawlak
Keyword(s):  
Signaling Pathway ◽  
Leucine Zipper ◽  
Site Directed Mutagenesis ◽  
3T3 Cells ◽  
Serum Response Element ◽  
Response Element ◽  
Nih 3T3 ◽  
Serum Response ◽  
Nih 3T3 Cells

Serum response element binding protein (SRE BP) is a novel binding factor present in nuclear extracts of avian and NIH 3T3 fibroblasts which specifically binds to the cfos SRE within a region overlapping and immediately 3' to the CArG box. Site-directed mutagenesis combined with transfection experiments in NIH 3T3 cells showed that binding of both serum response factor (SRF) and SRE BP is necessary for maximal serum induction of the SRE. In this study, we have combined size fractionation of the SRE BP DNA binding activity with C/EBPbeta antibodies to demonstrate that homodimers and heterodimers of p35C/EBPbeta (a transactivator) and p20C/EBPbeta (a repressor) contribute to the SRE BP complex in NIH 3T3 cells. Transactivation of the SRE by p35C/EBPbeta is dependent on SRF binding but not ternary complex factor (TCF) formation. Both p35C/EBPbeta and p20C/EBPbeta bind to SRF in vitro via a carboxy-terminal domain that probably does not include the leucine zipper. Moreover, SRE mutants which retain responsiveness to the TCF-independent signaling pathway bind SRE BP in vitro with affinities that are nearly identical to that of the wild-type SRE, whereas mutant SRE.M, which is not responsive to the TCF-independent pathway, has a nearly 10-fold lower affinity for SRE BP. We propose that C/EBPbeta may play a role in conjunction with SRF in the TCF-independent signaling pathway for SRE activation.

B-raf, a new member of the raf family, is activated by DNA rearrangement

1988 ◽  
Vol 8 (6) ◽  
pp. 2651-2654
Author(s):  
S Ikawa ◽  
M Fukui ◽  
Y Ueyama ◽  
N Tamaoki ◽  
T Yamamoto ◽  
...  
Keyword(s):  
Ewing Sarcoma ◽  
Dna Rearrangement ◽  
Terminal Portion ◽  
3T3 Cells ◽  
Complementary Dna ◽  
Amino Terminal ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Transforming Gene ◽  
Nih 3T3 Cells

Complementary DNA clones of a putative transforming gene were isolated from NIH 3T3 cells transformed with human Ewing sarcoma DNA. The gene was termed B-raf because it is related to but distinct from c-raf and A-raf. It appears that substitution in the amino-terminal portion of the normal B-raf protein confers transforming activity to the gene.

In vitro methylation of the human Ha-ras oncogene (T24) affects its transforming activity on NIH/3T3 cells

1986 ◽  
Vol 10 (3) ◽  
pp. 175-175
Author(s):  
M BORRELLO ◽  
M PIEROTTI ◽  
R DONGHI ◽  
I BONGARZONE ◽  
P MONDELLINI ◽  
...  
Keyword(s):  
Ras Oncogene ◽  
3T3 Cells ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

mos gene transforming efficiencies correlate with oocyte maturation and cytostatic factor activities

1991 ◽  
Vol 11 (2) ◽  
pp. 604-610
Author(s):  
N Yew ◽  
M Oskarsson ◽  
I Daar ◽  
D G Blair ◽  
G F Vande Woude
Keyword(s):  
Oocyte Maturation ◽  
Xenopus Oocytes ◽  
Kinase Activity ◽  
Vertebrate Species ◽  
3T3 Cells ◽  
Transforming Activity ◽  
Cytostatic Factor ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

The mos proto-oncogenes from different vertebrate species transform mouse NIH 3T3 cells with markedly different efficiencies. v-mos, mouse (c-mosmu), and chicken (c-mosch) mos transform NIH 3T3 cells 10- to 100-fold more efficiently than do human (c-moshu) and Xenopus (c-mosxc) mos. The mos genes with the highest transforming activity efficiently induce maturation in Xenopus oocytes and mimic cytostatic factor (CSF) by causing mitotic cleavage arrest in embryos. Chimeric v-mos/c-moshu proteins that had high transforming efficiencies in NIH 3T3 cells were also effective in the induction of oocyte maturation and CSF cleavage arrest. We measured the in vitro autophosphorylation activities of the different mos proteins and found that the levels of kinase activity of v-mos, c-mosmu, and c-mosch were much higher than that of c-mosxc. These data indicate that mos gene transforming efficiency and the ability to induce oocyte maturation or mimic CSF activity are correlated with in vitro autophosphorylation activity and suggest that the mos protein plays a similar role in transformed cells and normal oocytes.

scholarly journals RhoGEF Specificity Mutants Implicate RhoA as a Target for Dbs Transforming Activity

2002 ◽  
Vol 22 (19) ◽  
pp. 6895-6905 ◽  
Author(s):  
Li Cheng ◽  
Kent L. Rossman ◽  
Gwendolyn M. Mahon ◽  
David K. Worthylake ◽  
Malgorzata Korus ◽  
...  
Keyword(s):  
Cell Types ◽  
Competitive Inhibitor ◽  
Nucleotide Exchange ◽  
3T3 Cells ◽  
Structural Determinants ◽  
Nucleotide Exchange Factor ◽  
Transforming Activity ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

ABSTRACT Dbs is a Rho-specific guanine nucleotide exchange factor (RhoGEF) that exhibits transforming activity when overexpressed in NIH 3T3 mouse fibroblasts. Like many RhoGEFs, the in vitro catalytic activity of Dbs is not limited to a single substrate. It can catalyze the exchange of GDP for GTP on RhoA and Cdc42, both of which are expressed in most cell types. This lack of substrate specificity, which is relatively common among members of the RhoGEF family, complicates efforts to determine the molecular basis of their transforming activity. We have recently determined crystal structures of several RhoGEFs bound to their cognate GTPases and have used these complexes to predict structural determinants dictating the specificities of coupling between RhoGEFs and GTPases. Guided by this information, we mutated Dbs to alter significantly its relative exchange activity for RhoA versus Cdc42 and show that the transformation potential of Dbs correlates with exchange on RhoA but not Cdc42. Supporting this conclusion, oncogenic Dbs activates endogenous RhoA but not endogenous Cdc42 in NIH 3T3 cells. Similarly, a competitive inhibitor that blocks RhoA activation also blocks Dbs-mediated transformation. In conclusion, this study highlights the usefulness of specificity mutants of RhoGEFs as tools to genetically dissect the multiple signaling pathways potentially activated by overexpressed or oncogenic RhoGEFs. These ideas are exemplified for Dbs, which is strongly implicated in the transformation of NIH 3T3 cells via RhoA and not Cdc42.

Specific activation of the cellular Harvey-ras oncogene in dimethylbenzanthracene-induced mouse mammary tumors

1986 ◽  
Vol 6 (11) ◽  
pp. 4104-4108
Author(s):  
S Dandekar ◽  
S Sukumar ◽  
H Zarbl ◽  
L J Young ◽  
R D Cardiff
Keyword(s):  
Mammary Tumors ◽  
3T3 Cells ◽  
Mouse Mammary Tumor ◽  
Mouse Mammary Tumors ◽  
Transforming Activity ◽  
Radiation Induced ◽  
Genomic Dnas ◽  
Nih 3T3 ◽  
Adenosine Nucleotide ◽  
Nih 3T3 Cells

Genomic DNAs from dimethylbenzanthracene-induced BALB/c mouse mammary tumors arising from the transplantable hyperplastic outgrowth (HPO) line designated DI/UCD transformed NIH 3T3 cells upon transfection. Transforming activity was attributed to the presence of activated Harvey ras-1 oncogenes containing an A----T transversion at the middle adenosine nucleotide in codon 61. DNAs from untreated DI/UCD HPO cells and radiation-induced and spontaneous mammary tumors from the DI/UCD HPO line failed to transform NIH 3T3 cells. The results indicated that the mutation activation of Harvey ras-1 oncogenes was specific to dimethylbenzanthracene treatment in the mouse mammary tumor system.

scholarly journals The Role of Coconut Oil to Increase Expression of MMP-9, PDGF-BB, and TGF-β1 in NIH-3T3 Cell Line

2019 ◽  
Vol 7 (22) ◽  
pp. 3733-3736
Author(s):  
Dian Ika Perbina Meliala ◽  
Jansen Silalahi ◽  
Yuandani Yuandani ◽  
Linda Margata ◽  
Denny Satria
Keyword(s):  
Healing Process ◽  
Coconut Oil ◽  
Wound Healing Process ◽  
3T3 Cells ◽  
Virgin Coconut Oil ◽  
Nih3t3 Cells ◽  
Nih 3T3 ◽  
Tgf Β1 ◽  
Nih 3T3 Cells

AIM: The objective of the study was to evaluate protein expression in NIH 3T3 cells that are treated with virgin coconut oil (VCO) and hydrolysed of virgin coconut oil (HVCO) in vitro. METHODS: Coconut oil used in this study was virgin coconut oil (VCO) and VCO hydrolysed by Rhizomucor miehei (HVCO). NIH 3T3 cells (5x105 cells/well) were seeded in nine wells and incubated for overnight, then divided into three groups. Each group consisted of three wells. Group one without treatment, group two added VCO, and group three added HVCO and then incubated for overnight. One well in each group was added MMP-9, PDGF-BB, and TGF-β1 and incubated one hour. Finally, expressions of MMP-9, PDGF-BB, and TGF-β1 were detected using immunocytochemistry method. RESULTS: The results of the study showed that VCO and HVCO increased protein expressions of MMP-9, PDGF-BB, and TGF-β1. Percentage of MMP-9 expressions treated by VCO increased from 2.89 ± 0.07 to 28.16 ± 0.34, PDGF-BB from 28.11 ± 0.13 to 48.53 ± 0.49, and TGF-β1 from 4.19 ± 0.08 to 18.41 ± 0.54. Percentage of MMP-9 expressions treated by HVCO increased from 2.89 ± 0.07 to 55.40 ± 0.94, PDGF-BB from 28.11 ± 0.13 to 61.65 ± 0.42, and TGF-β1 from 4.19 ± 0.08 to 36.35 ± 0.67. CONCLUSION: VCO and HVCO increase the expression of MMP-9, PDGF-BB, dan TGF-β1 in NIH3T3 cells and therefore, coconut oil active in the wound healing process. HVCO is more than active than VCO.

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