Are oestrogen receptors and protein tyrosine kinases involved in phytoestrogen-modulated steroid secretion by porcine adrenocortical cells?

2012 ◽  
Vol 60 (2) ◽  
pp. 285-295
Author(s):  
Barbara Kaminska ◽  
Marek Opalka ◽  
Renata Ciereszko ◽  
Luiza Dusza
Keyword(s):  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Oestrous Cycle ◽  
Protein Tyrosine Kinases ◽  
Cortisol Secretion ◽  
Oestrogen Receptors ◽  
Observable Effect ◽  
Adrenocortical Cells ◽  
Protein Tyrosine ◽  
Induced Changes

The phytoestrogens genistein and daidzein had been found to affect the function of some tissues via oestrogen receptors (ER). In addition, genistein, but not daidzein, is considered to be a protein tyrosine kinase (PTK) inhibitor. Thus, the involvement of oestrogen receptors and PTK in phytoestrogen action on adrenocortical porcine steroidogenesis was examined in this study. The aims of the experiment were to test the effects of (i) ICI 182, 780 (ICI), an ER antagonist, on genistein- and daidzein-modulated cortisol and androstenedione (A4) secretion by adrenocortical cells isolated during the luteal and follicular phases of the porcine oestrous cycle; (ii) tyrphostin AG 957 (TAG), a nonsteroidal PTK inhibitor, on cortisol and A4 secretion by the cells and (iii) the phase of the porcine oestrous cycle on the mechanism of phytoestrogen action. Adrenals were harvested during the luteal (n = 5 animals) and follicular (n = 5 animals) phases of the oestrous cycle from locally slaughtered crossbred gilts. The isolated adrenocortical cells were incubated for 8 h (37 °C, 95% air, 5% CO2) with genistein (5 or 10 μM) or daidzein (5 or 10 μM) in the presence or absence of ICI (0.5 μM) or TAG (5 or 10 μM). Genistein and daidzein inhibited cortisol secretion and stimulated A4 secretion by porcine adrenocortical cells harvested during both the luteal and follicular phases of the oestrous cycle. The ER antagonist ICI did not eliminate phytoestrogen-induced changes in steroidogenesis. In contrast to genistein, TAG reduced the secretion of A4 and did not affect cortisol secretion. There was no observable effect due to the phase of the cycle. It is suggested that the mechanism of genistein and daidzein action in the adrenocortical cells of pigs is independent of ER and PTK. It is possible that PTK are involved in A4 secretion by porcine adrenocortical cells.

The myristylation signal of p60v-src functionally complements the N-terminal fps-specific region of P130gag-fps

1989 ◽  
Vol 9 (5) ◽  
pp. 2214-2219
Author(s):  
A R Brooks-Wilson ◽  
E Ball ◽  
T Pawson
Keyword(s):  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Specific Region ◽  
Protein Tyrosine Kinases ◽  
Membrane Association ◽  
Modular Construction ◽  
Specific Domain ◽  
Protein Tyrosine ◽  
Sarcoma Virus ◽  
Transforming Activity

The P130gag-fps protein-tyrosine kinase of Fujinami sarcoma virus contains an N-terminal fps-specific domain (Nfps) that is important for oncogenicity. The N-terminal 14 amino acids of p60v-src, which direct myristylation and membrane association, can replace the gag-Nfps sequences of P130gag-fps (residues 1 to 635), producing a highly transforming src-fps polypeptide. Conversely, gag-Nfps can restore modest transforming activity to a nonmyristylated v-src polypeptide. These results emphasize the modular construction of protein-tyrosine kinases and indicate that Nfps, possibly in conjunction with gag, functions in the subcellular localization of P130gag-fps.

Stable association of pp60src and pp59fyn with the focal adhesion-associated protein tyrosine kinase, pp125FAK

1994 ◽  
Vol 14 (1) ◽  
pp. 147-155
Author(s):  
B S Cobb ◽  
M D Schaller ◽  
T H Leu ◽  
J T Parsons
Keyword(s):  
Focal Adhesion ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Src Family Kinases ◽  
Protein Tyrosine Kinases ◽  
Protein Tyrosine ◽  
Sh2 Domains ◽  
Embryo Cells ◽  
Stable Association

Changes in cellular growth and dramatic alterations in cell morphology and adhesion are common features of cells transformed by oncogenic protein tyrosine kinases, such as pp60src and other members of the Src family. In this report, we present evidence for the stable association of two Src family kinases (pp60src and pp59fyn) with tyrosine-phosphorylated forms of a focal adhesion-associated protein tyrosine kinase, pp125FAK. In Src-transformed chicken embryo cells, most of the pp125FAK was stably complexed with activated pp60src (e.g., pp60(527F). The stable association of pp125FAK with pp60(527F) in vivo required the structural integrity of the Src SH2 domain. The association of pp60(527F) and pp125FAK could be reconstituted in vitro by incubation of normal cell extracts with glutathione S-transferase fusion proteins containing SH2 or SH3/SH2 domains of pp60src. Furthermore, the association of isolated SH2 or SH3/SH2 domains with in vitro 32P-labeled pp125FAK protected the major site of pp125FAK autophosphorylation from digestion with a tyrosine phosphatase, indicating that the autophosphorylation site of pp125FAK participates in binding with Src. Immunoprecipitation of Src family kinases from extracts of normal chicken embryo cells revealed stable complexes of pp59fyn and tyrosine-phosphorylated pp125FAK. These data provide evidence for a direct interaction between two cytoplasmic nonreceptor protein tyrosine kinases and suggest that Src may contribute to changes in pp125FAK regulation in transformed cells. Furthermore, pp125FAK may directly participate in the targeting of pp59fyn or possibly other Src family kinases to focal adhesions in normal cells.

scholarly journals Novel yeast protein kinase (YPK1 gene product) is a 40-kilodalton phosphotyrosyl protein associated with protein-tyrosine kinase activity.

1990 ◽  
Vol 10 (12) ◽  
pp. 6244-6256 ◽  
Author(s):  
D Dailey ◽  
G L Schieven ◽  
M Y Lim ◽  
H Marquardt ◽  
T Gilmore ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Kinase Activity ◽  
Protein Tyrosine Kinases ◽  
Tyrosine Kinase Activity ◽  
Protein Tyrosine ◽  
Protein Tyrosine Kinase Activity

Extracts of bakers' yeast (Saccharomyces cerevisiae) contain protein-tyrosine kinase activity that can be detected with a synthetic Glu-Tyr copolymer as substrate (G. Schieven, J. Thorner, and G.S. Martin, Science 231:390-393, 1986). By using this assay in conjunction with ion-exchange and affinity chromatography, a soluble tyrosine kinase activity was purified over 8,000-fold from yeast extracts. The purified activity did not utilize typical substrates for mammalian protein-tyrosine kinases (enolase, casein, and histones). The level of tyrosine kinase activity at all steps of each preparation correlated with the content of a 40-kDa protein (p40). Upon incubation of the most highly purified fractions with Mn-ATP or Mg-ATP, p40 was the only protein phosphorylated on tyrosine. Immunoblotting of purified p40 or total yeast extracts with antiphosphotyrosine antibodies and phosphoamino acid analysis of 32P-labeled yeast proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the 40-kDa protein is normally phosphorylated at tyrosine in vivo. 32P-labeled p40 immunoprecipitated from extracts of metabolically labeled cells by affinity-purified anti-p40 antibodies contained both phosphoserine and phosphotyrosine. The gene encoding p40 (YPK1) was cloned from a yeast genomic library by using oligonucleotide probes designed on the basis of the sequence of purified peptides. As deduced from the nucleotide sequence of YPK1, p40 is homologous to known protein kinases, with features that resemble known protein-serine kinases more than known protein-tyrosine kinases. Thus, p40 is a protein kinase which is phosphorylated in vivo and in vitro at both tyrosine and serine residues; it may be a novel type of autophosphorylating tyrosine kinase, a bifunctional (serine/tyrosine-specific) protein kinase, or a serine kinase that is a substrate for an associated tyrosine kinase.

scholarly journals Tec protein-tyrosine kinase is involved in interleukin-3 signaling pathway

Blood ◽  
1995 ◽  
Vol 85 (2) ◽  
pp. 343-350 ◽  
Author(s):  
H Mano ◽  
Y Yamashita ◽  
K Sato ◽  
Y Yazaki ◽  
H Hirai
Keyword(s):  
Signaling Pathway ◽  
Cell Lines ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Kinase Activity ◽  
Protein Tyrosine Kinases ◽  
Protein Tyrosine ◽  
Interleukin 3 ◽  
Hematopoietic Cell Lines

Abstract Among cytoplasmic protein-tyrosine kinases (PTKs) Tec now forms a novel subfamily with recently identified Tec-related PTKs (Btk and Itk/Tsk). Tec is known to be abundantly expressed in myeloid cells, and multiple forms of Tec protein can be generated via the mechanism of alternative splicing. In this report, we have investigated 5′-terminal diversity of the tec messages to demonstrate a predominant form of the Tec protein in mouse hematopoietic cell lines. Using anti-Tec serum, we could show that stimulation with interleukin-3 (IL-3) can induce tyrosine phosphorylation of Tec both in myeloid and pro-B-cell lines. IL-3 stimulation was also shown to induce kinase activity of Tec. Furthermore, we could demonstrate that Tec is constitutively associated with the Shc protein in vivo. Thus, we conclude that Tec is involved in the signaling pathway of IL-3.

Novel yeast protein kinase (YPK1 gene product) is a 40-kilodalton phosphotyrosyl protein associated with protein-tyrosine kinase activity

1990 ◽  
Vol 10 (12) ◽  
pp. 6244-6256
Author(s):  
D Dailey ◽  
G L Schieven ◽  
M Y Lim ◽  
H Marquardt ◽  
T Gilmore ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Kinase Activity ◽  
Protein Tyrosine Kinases ◽  
Tyrosine Kinase Activity ◽  
Protein Tyrosine ◽  
Protein Tyrosine Kinase Activity

Extracts of bakers' yeast (Saccharomyces cerevisiae) contain protein-tyrosine kinase activity that can be detected with a synthetic Glu-Tyr copolymer as substrate (G. Schieven, J. Thorner, and G.S. Martin, Science 231:390-393, 1986). By using this assay in conjunction with ion-exchange and affinity chromatography, a soluble tyrosine kinase activity was purified over 8,000-fold from yeast extracts. The purified activity did not utilize typical substrates for mammalian protein-tyrosine kinases (enolase, casein, and histones). The level of tyrosine kinase activity at all steps of each preparation correlated with the content of a 40-kDa protein (p40). Upon incubation of the most highly purified fractions with Mn-ATP or Mg-ATP, p40 was the only protein phosphorylated on tyrosine. Immunoblotting of purified p40 or total yeast extracts with antiphosphotyrosine antibodies and phosphoamino acid analysis of 32P-labeled yeast proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the 40-kDa protein is normally phosphorylated at tyrosine in vivo. 32P-labeled p40 immunoprecipitated from extracts of metabolically labeled cells by affinity-purified anti-p40 antibodies contained both phosphoserine and phosphotyrosine. The gene encoding p40 (YPK1) was cloned from a yeast genomic library by using oligonucleotide probes designed on the basis of the sequence of purified peptides. As deduced from the nucleotide sequence of YPK1, p40 is homologous to known protein kinases, with features that resemble known protein-serine kinases more than known protein-tyrosine kinases. Thus, p40 is a protein kinase which is phosphorylated in vivo and in vitro at both tyrosine and serine residues; it may be a novel type of autophosphorylating tyrosine kinase, a bifunctional (serine/tyrosine-specific) protein kinase, or a serine kinase that is a substrate for an associated tyrosine kinase.

scholarly journals Cytosolic Protein-Tyrosine Kinase Activities in Various Rat Tissues

1989 ◽  
Vol 26 (2) ◽  
pp. 164-168 ◽  
Author(s):  
Tomoko Kobayashi ◽  
Shun-Ichi Nakamura ◽  
Hirohei Yamamura
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Total Activity ◽  
Protein Tyrosine Kinases ◽  
Rat Tissues ◽  
Protein Tyrosine ◽  
Cytosolic Protein ◽  
Lymphatic Organs ◽  
Assay Conditions

Suitable assay conditions for the detection of cytosolic protein-tyrosine kinase activities in crude extracts of various rat tissues have been determined. Cytosolic protein-tyrosine kinases showed common characteristics including substrate specificity and divalent cation requirement. Using (Val5) angiotensin II and Mn2+ rather than a src-related synthetic peptide, E11G1, and Mg2+, we obtained higher activities of cytosolic protein-tyrosine kinases. Among various rat tissues tested, spleen, bone marrow, thymus, small intestine, appendix and lung, in decreasing order of total activity, contained high activities of cytosolic protein-tyrosine kinases. These results suggest that the enzyme activities in lymphatic organs and in organs closely related to cell proliferation are high. The assay system described allows the precise measurement of cytosolic protein-tyrosine kinase activity in various rat tissues, both normal and malignant.

scholarly journals Differential modulation of plasminogen activator gene expression by oncogene-encoded protein tyrosine kinases.

1993 ◽  
Vol 13 (9) ◽  
pp. 5888-5897 ◽  
Author(s):  
S M Bell ◽  
D C Connolly ◽  
N J Maihle ◽  
J L Degen
Keyword(s):  
Gene Expression ◽  
Protein Kinase ◽  
Tyrosine Kinase ◽  
Plasminogen Activator ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Protein Tyrosine Kinases ◽  
Mrna Levels ◽  
Protein Tyrosine ◽  
Upa Mrna

Urokinase-type plasminogen activator (uPA) gene transcription is increased > or = 50-fold in chicken embryo fibroblasts (CEF) following transformation by the protein tyrosine kinase pp60v-src. Protein phosphorylation appears to play a critical role in uPA gene expression in these cells; protein kinase C-activating phorbol esters cooperate with pp60v-src to synergistically increase uPA mRNA, whereas cyclic AMP (cAMP)-dependent protein kinase-activating agents (e.g., 8-bromo cAMP) repress uPA mRNA levels. To explore the relationship between transforming oncogenes and uPA gene expression, uPA mRNA levels were measured in CEF infected with selected avian retroviruses. We report that v-ras and the transforming protein tyrosine kinases v-src, v-yes, and v-ros all increase cellular uPA mRNAs. However, transformation with the protein tyrosine kinase encoded by v-erbB, or the nuclear proteins encoded by v-jun, v-ski, or v-myc, did not increase uPA mRNA detectably. Ras and all of the protein tyrosine kinases analyzed, including the v-erbB product, but none of the nuclear oncoproteins sensitized cells to phorbol ester induction of uPA gene expression. Thus, increased uPA gene expression is not simply a secondary consequence of cell transformation but, rather, is regulated or comodulated by only a subset of oncogene products. Analysis of cells expressing site-directed mutants of pp60v-src showed that the induction of the uPA gene is dependent on protein tyrosine kinase catalytic activity, myristylation, and plasma membrane localization. However, these properties together are not sufficient; an additional feature in the src homology 2 domain is also required. The major sites of serine phosphorylation, serines 12 and 17, and the autophosphorylation site, tyrosine 416, are not essential for uPA gene induction. However, the reduction of uPA mRNA in pp60v-src-transformed cells by 8-bromo cAMP is dependent on tyrosine 416.

scholarly journals Augmented expression of a myeloid-specific protein tyrosine kinase gene (hck) after macrophage activation.

1988 ◽  
Vol 168 (5) ◽  
pp. 1801-1810 ◽  
Author(s):  
S F Ziegler ◽  
C B Wilson ◽  
R M Perlmutter
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Cell Activity ◽  
Myeloid Cell ◽  
Specific Protein ◽  
Protein Tyrosine Kinases ◽  
Kinase Gene ◽  
Protein Tyrosine ◽  
Family Protein

Protein tyrosine kinases are thought to participate in signal transduction pathways in a variety of cell types. Recent studies have identified a new src family protein tyrosine kinase (hck) that is preferentially expressed in myeloid cells. To examine the hypothesis that this kinase may regulate myeloid cell activity, antisera were generated that define the 59-kD product of the hck gene. Functional activation of human cultured macrophages with LPS augmented the expression of hck transcripts and of p59hck, but decreased the level of transcripts encoded by the closely related c-fgr protooncogene. Thus these two structurally similar src family kinases almost certainly subserve distinct functions. Reasoning from the known properties of the src family protein tyrosine kinases, it is likely that the products of these two protooncogenes assist in regulating the behavior of activated phagocytes.

Thrombin-dependent association of phosphatidylinositol-3 kinase with p60c-src and p59fyn in human platelets

1990 ◽  
Vol 10 (7) ◽  
pp. 3806-3809
Author(s):  
J S Gutkind ◽  
P M Lacal ◽  
K C Robbins
Keyword(s):  
Tyrosine Kinases ◽  
Protein Tyrosine Kinase ◽  
Human Platelets ◽  
Protein Tyrosine Kinases ◽  
Reaction Products ◽  
Phosphatidylinositol 3 Kinase ◽  
Protein Tyrosine ◽  
Nonreceptor Protein Tyrosine Kinases ◽  
Pi Kinase ◽  
Phosphatidylinositol 3

Recent studies have shown that ligand-activated growth factor receptors as well as transforming versions of nonreceptor protein-tyrosine kinases physically associate with phosphatidylinositol-3 kinase (PI-3 kinase). Reasoning that PI-3 kinase might also play a role in the normal functions of nonreceptor kinases, we sought to determine whether association with PI-3 kinase might serve as a measure of nonreceptor protein-tyrosine kinase activation under physiological conditions. We found that p60c-src as well as p59fyn, the product of another member of the src family of proto-oncogenes, physically associated with a PI kinase activity within 5 s after exposure to thrombin. Furthermore, PI kinase reaction products generated in p60v-src, p60c-src or p59fyn containing immunoprecipitates were indistinguishable, demonstrating the identity of the associated enzyme as PI-3 kinase. These findings demonstrate a thrombin-dependent interaction between p60c-src or p59fyn and PI-3 kinase and suggest a role for nonreceptor protein-tyrosine kinases in human platelet signal transduction.

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