Analysis of phorbol ester receptors in phorbol ester unresponsive 3T3 cell variants

Blumberg, Peter M.; Butler‐Gralla, Edith; Herschman, Harvey R.

doi:10.1016/0006-291x(81)91611-9

Cited by 29 publications

(11 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This is consistent with earlier observations that 3T3-TNR9 cells possess a phorbol ester binding on May 10, 2018 by guest http://mcb.asm.org/ Downloaded from activity and are able to respond to TPA with increases in glucose transport and arachidonic acid release (6,7,9). Thus, a lesion in some postreceptor step in the sequence of events leading to mitogenesis is likely to be responsible for the failure of 3T3-TNR9 cells to respond mitogenically to TPA.…”

Section: Discussionsupporting

confidence: 92%

“…cells, 3T3-TNR9, cannot be stimulated to enter the DNAsynthetic phase of the cell cycle in response to TPA, but retains the ability to reenter the cell cycle in response to FGF, EGF, or serum (7). Binding studies with whole cells, using isotopically labeled phorbol dibutyrate, suggested that the 3T3-TNR9 variant retains TPA "receptors" (6). TPA stimulates several aspects of the pleiotypic response in the nonproliferative variant, including glucose transport and arachidonic acid release, but is unable to stimulate, in the 3T3-TNR9 cells, the elevation of ornithine decarboxylase seen in the parental cells (8,9).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

Bishop¹,

Martinez²,

Weber³

et al. 1985

Mol. Cell. Biol.

View full text Add to dashboard Cite

The 3T3-TNR9 cell line is a variant of Swiss 3T3 cells which does not respond mitogenically to tumor promoters, but does respond mitogenically to epidermal growth factor, fibroblast growth factor, and serum. To elucidate differences between tumor promoters and polypeptide mitogens in the pathway(s) of mitogenesis which might be responsible for the nonresponsiveness of the 3T3-TNR9 cells, we have examined in these cells the early protein phosphorylation events known to be associated with mitogenesis in the parental 3T3 cells. We find that the 3T3-TNR9 cells display levels of tetradecanoyl phorbol acetate binding and of a calcium- and phospholipid-dependent protein kinase activity which are at least the equal of those seen in the parental 3T3 cells, implicating some postreceptor event in the nonmitogenic phenotype. In addition, we find that phosphorylation of the epidermal growth factor receptor and of 80-kDa and 22-kDa proteins, as well as the tyrosine phosphorylation of a 42-kDa protein, all proceed normally in the nonmitogenic variant, even though these phosphorylations must depend on the activation of different kinases. Thus, all these early phosphorylation reactions are intact in the 3T3-TNR9 cells. Although these phosphorylations may be necessary, they clearly are insufficient to trigger mitogenesis.

show abstract

Section: Discussionsupporting

confidence: 92%

mentioning

confidence: 99%

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

Bishop¹,

Martinez²,

Weber³

et al. 1985

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

“…3T3-TNR9 cells contain normal levels of functional PKC (6), and many of the early responses to TPA which occur in the parental 3T3 cells also occur in the variant, including phosphorylation of p80 and p22 (5; but see reference 3) and increased prostaglandin biosynthesis (10) and glucose transport (9). Furthermore, several genes that are specifically inducible by TPA in Swiss 3T3 cells are also expressed in the same inducible manner in 3T3-TNR9 cells (31).…”

Section: Methodsmentioning

confidence: 99%

A Swiss 3T3 variant cell line resistant to the effects of tumor promoters cannot be transformed by src.

Nori¹,

Shawver²,

Weber³

1990

Mol. Cell. Biol.

View full text Add to dashboard Cite

To study the relationship between oncogenesis by v-src and normal cellular signalling pathways, we determined the effects of v-src on 3T3-TNR9 cells, a Swiss 3T3 variant which does not respond mitogenically to tumor promoters such as 12-O-tetradecanoyl-phorbol-13-acetate (TPA). We found that src was unable to transform these variant cells, whether the oncogene was introduced by infection with a murine retrovirus vector or by transfection with plasmid DNA. 3T3-TNR9 cells were not inherently resistant to transformation, since infection with similar recombinant retroviruses containing either v-ras or v-abl did induce transformation. Further analysis of Swiss 3T3 and 3T3-TNR9 cell populations infected with the v-src-containing retrovirus revealed that although the amount of v-src DNA in each was approximately the same, the level of the v-src message and protein and the overall level of phosphotyrosine expressed in the infected variants was much less than in infected parental cells. Cotransfection experiments using separate v-src and neo plasmids revealed a decrease in the number of G418-resistant colonies when transfections of TNR9 cells occurred in the presence of the src-containing plasmid, suggesting a growth inhibitory effect of v-src on 3T3-TNR9 cells, as has also been found for TPA itself. Since v-src cannot transform this variant cell line, which does not respond mitogenically to the protein kinase C agonist TPA, we suggest that src makes use of the protein kinase C pathway as part of its signalling activities.

show abstract

“…In summary, our previous data demonstrating the presence of TPA receptors on the TPA nonproliferative variant cells (Blumberg et al, 1981) suggest that distinct biochemical events distal to mitogen binding to receptors must occur in response to different mitogens. The inability of these cells to induce substantial levels of ODC in response to TPA further supports the existence of distinct postreceptor events, initiated by distinct mitogens, capable of leading to DNA synthesis and cell division.…”

Section: Discussionmentioning

confidence: 91%

Glucose uptake and ornithine decarboxylase activity in tetradecanoyl phorbol acetate non‐proliferative variants

Butler‐Gralla

Herschman

1983

Journal Cellular Physiology

Self Cite

View full text Add to dashboard Cite

The potent tumor promoter 12-0-tetradecanoyl phorbol-13-acetate (TPA) is also an excellant mitogen for 3T3 cells. We have previously isolated two independent variants, 3T3-TNR-2 and 3T3-TNR-9, that are unable to divide in response to TPA (Butler-Gralla and Herschman, 1981). We have now tested two components of the pleiotypic response, elevation of 2-deoxyglucose uptake and ornithine decarboxylase induction, in these cells. Basal levels of 2-deoxyglucose uptake were nearly tenfold higher in confluent 3T3-TNR-2 and 3T3-TNR-9 cells than in 3T3 cells. In contrast, basal ornithine decarboxylase levels were five- to tenfold lower in the variants. TPA stimulation of 2-deoxyglucose uptake was as great in absolute terms in the variant cell lines as that of 3T3 cells but was only half that observed with serum. TPA was unable to induce any elevation of ornithine decarboxylase in 3T3-TNR-9 cells. Although an elevation of ornithine decarboxylase levels occurred in 3T3-TNR-2 cells treated with TPA, the maximal specific activity in the variant was less than the unstimulated value for 3T3 cells.

show abstract

Analysis of phorbol ester receptors in phorbol ester unresponsive 3T3 cell variants

Cited by 29 publications

References 11 publications

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

A Swiss 3T3 variant cell line resistant to the effects of tumor promoters cannot be transformed by src.

Glucose uptake and ornithine decarboxylase activity in tetradecanoyl phorbol acetate non‐proliferative variants

Contact Info

Product

Resources

About