The Experts below are selected from a list of 207 Experts worldwide ranked by ideXlab platform
Irina V. Ogneva - One of the best experts on this subject based on the ideXlab platform.
-
Relative contents of cytoskeletal proteins in the membrane (MF) and cytoplasmic (CF) fractions of cardiomyocytes with typical Western blot images.
2018Co-Authors: Irina V. Ogneva, Sergey S. Loktev, Vladimir N. SychevAbstract:(A) Alpha-Actinin-1. (B) Alpha-Actinin-4. (C) Beta-actin. (D) Gamma-actin. (E) Beta-tubulin. (F) Desmin. “B”–basal control group, “V”–vivarium control group, “G”–ground control group (level marked by dotted line), “F”–flight group. *–p < 0.05 in comparison with group “G”. The values used to build graphs represented in the S1 Table. There were no changes in the cytoskeletal proteins contents in the membrane fraction of the cardiac tissue. In the cytoplasmic fraction, Alpha-Actinin-1 and Alpha-Actinin-4 protein content decreased during space flight.
-
Relative contents of cytoskeletal proteins in the membrane (MF) and cytoplasmic (CF) fractions of lung cells with typical Western blot images.
2018Co-Authors: Irina V. Ogneva, Sergey S. Loktev, Vladimir N. SychevAbstract:(A) Alpha-Actinin-1. (B) Alpha-Actinin-4. (C) Beta-actin. (D) Gamma-actin. (E) Beta-tubulin. (F) Desmin. “B”–basal control group, “V”–vivarium control group, “G”–ground control group (level marked by dotted line), “F”–flight group. The values used to build graphs represented in the S2 Table. There were no changes of cytoskeletal proteins contents in the membrane and cytoplasmic fractions of the lung tissue.
-
Lecithin Prevents Cortical Cytoskeleton Reorganization in Rat Soleus Muscle Fibers under Short-Term Gravitational Disuse
PloS one, 2016Co-Authors: Irina V. Ogneva, Nikolay S. BiryukovAbstract:The aim of this study was to prevent the cortical cytoskeleton reorganization of rat soleus muscle fibers under short-term gravitational disuse. Once a day, we injected the right soleus muscle with 0.5 ml lecithin at a concentration of 200 mg/ml and the left soleus muscle with a diluted solution in an equal volume for 3 days prior to the experiment. To simulate microgravity conditions in rats, an anti-orthostatic suspension was used according to the Ilyin-Novikov method modified by Morey-Holton et al. for 6 hours. The following groups of soleus muscle tissues were examined: "C", "C+L", "HS", and "HS+L". The transversal stiffness of rat soleus muscle fibers after 6 hours of suspension did not differ from that of the control group for the corresponding legs; there were no differences between the groups without lecithin «C» and «HS» or between the groups with lecithin "C+L" and "HS+L". However, lecithin treatment for three days resulted in an increase in cell stiffness; in the "C+L" group, cell stiffness was significantly higher by 22.7% (p < 0.05) compared with that of group "C". The mRNA content of genes encoding beta- and gamma-actin and beta-tubulin did not significantly differ before and after suspension in the corresponding groups. However, there was a significant increase in the mRNA content of these genes after lecithin treatment: the beta-actin and gamma-actin mRNA content in group "C+L" increased by 200% compared with that of group "C", and beta-tubulin increased by 100% (as well as the mRNA content of tubulin-binding proteins Ckap5, Tcp1, Cct5 and Cct7). In addition, desmin mRNA content remained unchanged in all of the experimental groups. As a result of the lecithin injections, there was a redistribution of the mRNA content of genes encoding actin monomer- and filament-binding proteins in the direction of increasing actin polymerization and filament stability; the mRNA content of Arpc3 and Lcp1 increased by 3- and 5-fold, respectively, but the levels of Tmod1 and Svil decreased by 2- and 5-fold, respectively. However, gravitational disuse did not result in changes in the mRNA content of Arpc3, Tmod1, Svil or Lcp1. Anti-orthostatic suspension for 6 hours resulted in a decrease in the mRNA content of Alpha-Actinin-4 (Actn4) and Alpha-Actinin-1 (Actn1) in group "HS" compared with that of group "C" by 25% and 30%, respectively, as well as a decrease and increase in the ACTN4 protein content in the membrane and cytoplasmic fractions, respectively. Lecithin injection resulted in an increase in the Actn1 and Actn4 mRNA content in group "C+L" by 1.5-fold and more than 2-fold, respectively, compared with the levels in group "C". Moreover, in group "HS+L", the mRNA content did not change in these genes compared with the levels in group "C+L", and the ACTN4 protein content in the membrane and cytoplasmic fractions also remained unchanged. Thus, lecithin prevented the reduction of Actn1 and Actn4 mRNA and the migration of ACTN4 from the cortical cytoskeleton to the cytoplasm.
-
Possible role of non-muscle Alpha-Actinins in muscle cell mechanosensitivity.
PloS one, 2014Co-Authors: Irina V. Ogneva, Nikolay S. Biryukov, Toomas A. Leinsoo, Irina M. LarinaAbstract:The main hypothesis suggested that changes in the external mechanical load would lead to different deformations of the submembranous cytoskeleton and, as a result, dissociation of different proteins from its structure (induced by increased/decreased mechanical stress). The study subjects were fibers of the soleus muscle and cardiomyocytes of Wistar rats. Changes in external mechanical conditions were reconstructed by means of antiorthostatic suspension of the animals by their tails for 6, 12, 18, 24 and 72 hours. Transversal stiffness was measured by atomic force microscopy imaging; beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 levels in membranous and cytoplasmic fractions were quantified by Western blot analysis; expression rates of the corresponding genes were studied using RT-PCR. Results: In 6 hours, Alpha-Actinin 1 and Alpha-Actinin 4 levels decreased in the membranous fraction of proteins of cardiomyocytes and soleus muscle fibers, respectively, but increased in the cytoplasmic fraction of the abovementioned cells. After 6–12 hours of suspension, the expression rates of beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 were elevated in the soleus muscle fibers, but the Alpha-Actinin 1 expression rate returned to the reference level in 72 hours. After 18–24 hours, the expression rates of beta-actin and Alpha-Actinin 4 increased in cardiomyocytes, while the Alpha-Actinin 1 expression rate decreased in soleus muscle fibers. After 12 hours, the beta- and gamma-actin content dropped in the membranous fraction and increased in the cytoplasmic protein fractions from both cardiomyocytes and soleus muscle fibers. The stiffness of both cell types decreased after the same period of time. Further, during the unloading period the concentration of nonmuscle actin and different isoforms of Alpha-Actinins increased in the membranous fraction from cardiomyocytes. At the same time, the concentration of the abovementioned proteins decreased in the soleus muscle fibers.
-
Structure of cortical cytoskeleton in fibers of mouse muscle cells after being exposed to a 30-day space flight on board the BION-M1 biosatellite.
Journal of applied physiology (Bethesda Md. : 1985), 2014Co-Authors: Irina V. Ogneva, Maria V. Maximova, Irina M. LarinaAbstract:The aim of the work was to analyze changes in the organization of the cortical cytoskeleton in fibers of the mouse soleus muscle, tibialis anterior muscle and left ventricular cardiomyocytes after completion of a 30-day space flight on board the BION-M1 biosatellite (Russia, 2013). The transversal stiffness of the cortical cytoskeleton of the cardiomyocytes and fibers of the skeletal muscles did not differ significantly within the study groups compared with the vivarium control group. The content of beta- and gamma-actin in the membranous fraction of proteins in the left ventricular cardiomyocytes did not differ significantly within all study groups and correlated with the transversal stiffness. A similar situation was revealed in fibers of the soleus muscle and tibialis anterior muscle. At the same time, the content of beta-actin in the cytoplasmic fraction of proteins was found to be decreased in all types of studied tissues compared with the control levels in the postflight group, with lowered beta-actin gene expression rates in the postflight group. After completion of the space flight, the content of Alpha-Actinin-4 was found to be reduced in the membranous fraction of proteins from the mouse cardiomyocytes, while its content in the cytoplasmic fraction of proteins did not change significantly. Furthermore, gene expression rates of this protein were decreased at the time of dissection (it was started after 13 h after landing). At the same time, the content of Alpha-Actinin-1 decreased in the membranous fraction and increased in the cytoplasmic fraction of proteins from the soleus muscle fibers.
Irina M. Larina - One of the best experts on this subject based on the ideXlab platform.
-
Proteomic profile of cultured human endothelial cells after exposition to simulated microgravity
Acta Astronautica, 2021Co-Authors: D. N. Kashirina, Irina M. Larina, Alexey S. Kononikhin, Andrey Ratushnyy, Evgeny N. Nikolaev, Ludmila BuravkovaAbstract:Abstract The proteome of human umbilical vein endothelial cells (HUVEC) cultured under static conditions and simulated microgravity (sμG) using Random Positioning Machine (RPM) for 24 h was studied by chromatography-mass spectrometry. It was revealed that the percentage of ribosomal proteins and proteins involved in intercellular adhesion, regulation of actin cytoskeleton, various pathways of cell signaling mediated by G-proteins, apoptosis and ubiquitin-dependent protein catabolism increased under sμG. At the same time the number of proteins associated with cell growth reduced. A significant increase in the peak intensities of proteins associated with maintaining the cytoskeleton and stress fibers (myristoylated alanine-rich C-kinase substrate, filamin-A, Alpha-Actinin-1 and myosin light polypeptide 6) and proteins involved in response to unfolded protein (serpin H1, 78 kDa glucose-regulated protein, transitional endoplasmic reticulum ATPase) was shown. At the same time, a significant decrease in the peak intensity of cofilin 1, which is capable of breaking actin filaments, was revealed. Thus, the most pronounced effect of microgravity is on the proteins associated with the actin cytoskeleton, as well as on adhesion proteins, whose properties also depend on the structure of the cytoskeleton. At the same time, the number of proteins which prevent improper protein folding increases, and the translation apparatus is rearranged under sμG.
-
Possible role of non-muscle Alpha-Actinins in muscle cell mechanosensitivity.
PloS one, 2014Co-Authors: Irina V. Ogneva, Nikolay S. Biryukov, Toomas A. Leinsoo, Irina M. LarinaAbstract:The main hypothesis suggested that changes in the external mechanical load would lead to different deformations of the submembranous cytoskeleton and, as a result, dissociation of different proteins from its structure (induced by increased/decreased mechanical stress). The study subjects were fibers of the soleus muscle and cardiomyocytes of Wistar rats. Changes in external mechanical conditions were reconstructed by means of antiorthostatic suspension of the animals by their tails for 6, 12, 18, 24 and 72 hours. Transversal stiffness was measured by atomic force microscopy imaging; beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 levels in membranous and cytoplasmic fractions were quantified by Western blot analysis; expression rates of the corresponding genes were studied using RT-PCR. Results: In 6 hours, Alpha-Actinin 1 and Alpha-Actinin 4 levels decreased in the membranous fraction of proteins of cardiomyocytes and soleus muscle fibers, respectively, but increased in the cytoplasmic fraction of the abovementioned cells. After 6–12 hours of suspension, the expression rates of beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 were elevated in the soleus muscle fibers, but the Alpha-Actinin 1 expression rate returned to the reference level in 72 hours. After 18–24 hours, the expression rates of beta-actin and Alpha-Actinin 4 increased in cardiomyocytes, while the Alpha-Actinin 1 expression rate decreased in soleus muscle fibers. After 12 hours, the beta- and gamma-actin content dropped in the membranous fraction and increased in the cytoplasmic protein fractions from both cardiomyocytes and soleus muscle fibers. The stiffness of both cell types decreased after the same period of time. Further, during the unloading period the concentration of nonmuscle actin and different isoforms of Alpha-Actinins increased in the membranous fraction from cardiomyocytes. At the same time, the concentration of the abovementioned proteins decreased in the soleus muscle fibers.
-
Structure of cortical cytoskeleton in fibers of mouse muscle cells after being exposed to a 30-day space flight on board the BION-M1 biosatellite.
Journal of applied physiology (Bethesda Md. : 1985), 2014Co-Authors: Irina V. Ogneva, Maria V. Maximova, Irina M. LarinaAbstract:The aim of the work was to analyze changes in the organization of the cortical cytoskeleton in fibers of the mouse soleus muscle, tibialis anterior muscle and left ventricular cardiomyocytes after completion of a 30-day space flight on board the BION-M1 biosatellite (Russia, 2013). The transversal stiffness of the cortical cytoskeleton of the cardiomyocytes and fibers of the skeletal muscles did not differ significantly within the study groups compared with the vivarium control group. The content of beta- and gamma-actin in the membranous fraction of proteins in the left ventricular cardiomyocytes did not differ significantly within all study groups and correlated with the transversal stiffness. A similar situation was revealed in fibers of the soleus muscle and tibialis anterior muscle. At the same time, the content of beta-actin in the cytoplasmic fraction of proteins was found to be decreased in all types of studied tissues compared with the control levels in the postflight group, with lowered beta-actin gene expression rates in the postflight group. After completion of the space flight, the content of Alpha-Actinin-4 was found to be reduced in the membranous fraction of proteins from the mouse cardiomyocytes, while its content in the cytoplasmic fraction of proteins did not change significantly. Furthermore, gene expression rates of this protein were decreased at the time of dissection (it was started after 13 h after landing). At the same time, the content of Alpha-Actinin-1 decreased in the membranous fraction and increased in the cytoplasmic fraction of proteins from the soleus muscle fibers.
-
Structure of the cortical cytoskeleton of the left cardiac ventricle hystiocytes and of soleus muscle fibers in rats following short-time suspension
Aviakosmicheskaia i ekologicheskaia meditsina = Aerospace and environmental medicine, 2014Co-Authors: Ogneva, Irina M. LarinaAbstract:Purpose of the investigation was to examine structural rearrangements in the cortical cytoskeleton as a result of changes in external mechanic conditions. Objects of the investigation were Wistar rat's m. soleus fibers and left ventricle hystiocytes. Suspension for 6, 12, 18, 24 and 72 hours was performed according to the Ilyin-Novikov procedure modified by Morey-Holton. Cell stiffness, non-muscle actin and actin-binding proteins in the protein membrane and cytoplasmic fractions, and respective gene expression were evaluated. In addition, corticosterone levels were measured in blood serum. After 6 hours of suspension, actin-binding proteins went down in the membrane fraction, i.e. Alpha-Actinin-1 decreased in hystiocytes and Alpha-Actinin-4 in m. soleus fibers. On the contrary, their content in the cytoplasmatic fraction increased. Expression of genes coding beta- and gamma-actin, Alpha-Actinin-1 and Alpha-Actinin-4 in m. soleus fibers showed a decrease. However, the apha-Actinin-1 gene regained its baseline expression rate in 72 hours. Following 18 to 24 hours of suspension, expression of the beta-actin and Alpha-Actinin-4 genes in hystiocytes grew in comparison with baseline values, while expression of Alpha-Actinin-1 gene decreased. After 12 hours of suspension, beta- and gamma-actin levels were reduced in membrane proteins and increased in cytoplasmatic proteins in hystiocytes and m. soleus fibers. Stiffness showed a decline in each type of cell. Further into suspension, membrane proteins of hystiocytes increased levels of non-muscle actin and actin-binding proteins, and stiffness of these cells. Levels of these proteins decreased in m. soleus fibers, as also did cell stiffness. Serum corticosterone hardly altered but was slightly increased after 6 hours of suspension.
-
Structure and functional characteristics of rat's left ventricle cardiomyocytes under antiorthostatic suspension of various duration and subsequent reloading.
Journal of biomedicine & biotechnology, 2012Co-Authors: Irina V. Ogneva, Nikolay S. Biryukov, T. M. Mirzoev, O. M. Veselova, Irina M. LarinaAbstract:The goal of the research was to identify the structural and functional characteristics of the rat's left ventricle under antiorthostatic suspension within 1, 3, 7 and 14 days, and subsequent 3 and 7-day reloading after a 14-day suspension. The transversal stiffness of the cardiomyocyte has been determined by the atomic force microscopy, cell respiration—by polarography and proteins content—by Western blotting. Stiffness of the cortical cytoskeleton increases as soon as one day after the suspension and increases up to the 14th day, and starts decreasing during reloading, reaching the control level after 7 days. The stiffness of the contractile apparatus and the intensity of cell respiration also increases. The content of non-muscle isoforms of actin in the cytoplasmic fraction of proteins does not change during the whole experiment, as does not the beta-actin content in the membrane fraction. The content of gamma-actin in the membrane fraction correlates with the change in the transversal stiffness of the cortical cytoskeleton. Increased content of Alpha-Actinin-1 and Alpha-Actinin-4 in the membrane fraction of proteins during the suspension is consistent with increased gamma-actin content there. The opposite direction of change of Alpha-Actinin-1 and Alpha-Actinin-4 content suggests their involvement into the signal pathways.
Nikolay S. Biryukov - One of the best experts on this subject based on the ideXlab platform.
-
Lecithin Prevents Cortical Cytoskeleton Reorganization in Rat Soleus Muscle Fibers under Short-Term Gravitational Disuse
PloS one, 2016Co-Authors: Irina V. Ogneva, Nikolay S. BiryukovAbstract:The aim of this study was to prevent the cortical cytoskeleton reorganization of rat soleus muscle fibers under short-term gravitational disuse. Once a day, we injected the right soleus muscle with 0.5 ml lecithin at a concentration of 200 mg/ml and the left soleus muscle with a diluted solution in an equal volume for 3 days prior to the experiment. To simulate microgravity conditions in rats, an anti-orthostatic suspension was used according to the Ilyin-Novikov method modified by Morey-Holton et al. for 6 hours. The following groups of soleus muscle tissues were examined: "C", "C+L", "HS", and "HS+L". The transversal stiffness of rat soleus muscle fibers after 6 hours of suspension did not differ from that of the control group for the corresponding legs; there were no differences between the groups without lecithin «C» and «HS» or between the groups with lecithin "C+L" and "HS+L". However, lecithin treatment for three days resulted in an increase in cell stiffness; in the "C+L" group, cell stiffness was significantly higher by 22.7% (p < 0.05) compared with that of group "C". The mRNA content of genes encoding beta- and gamma-actin and beta-tubulin did not significantly differ before and after suspension in the corresponding groups. However, there was a significant increase in the mRNA content of these genes after lecithin treatment: the beta-actin and gamma-actin mRNA content in group "C+L" increased by 200% compared with that of group "C", and beta-tubulin increased by 100% (as well as the mRNA content of tubulin-binding proteins Ckap5, Tcp1, Cct5 and Cct7). In addition, desmin mRNA content remained unchanged in all of the experimental groups. As a result of the lecithin injections, there was a redistribution of the mRNA content of genes encoding actin monomer- and filament-binding proteins in the direction of increasing actin polymerization and filament stability; the mRNA content of Arpc3 and Lcp1 increased by 3- and 5-fold, respectively, but the levels of Tmod1 and Svil decreased by 2- and 5-fold, respectively. However, gravitational disuse did not result in changes in the mRNA content of Arpc3, Tmod1, Svil or Lcp1. Anti-orthostatic suspension for 6 hours resulted in a decrease in the mRNA content of Alpha-Actinin-4 (Actn4) and Alpha-Actinin-1 (Actn1) in group "HS" compared with that of group "C" by 25% and 30%, respectively, as well as a decrease and increase in the ACTN4 protein content in the membrane and cytoplasmic fractions, respectively. Lecithin injection resulted in an increase in the Actn1 and Actn4 mRNA content in group "C+L" by 1.5-fold and more than 2-fold, respectively, compared with the levels in group "C". Moreover, in group "HS+L", the mRNA content did not change in these genes compared with the levels in group "C+L", and the ACTN4 protein content in the membrane and cytoplasmic fractions also remained unchanged. Thus, lecithin prevented the reduction of Actn1 and Actn4 mRNA and the migration of ACTN4 from the cortical cytoskeleton to the cytoplasm.
-
Possible role of non-muscle Alpha-Actinins in muscle cell mechanosensitivity.
PloS one, 2014Co-Authors: Irina V. Ogneva, Nikolay S. Biryukov, Toomas A. Leinsoo, Irina M. LarinaAbstract:The main hypothesis suggested that changes in the external mechanical load would lead to different deformations of the submembranous cytoskeleton and, as a result, dissociation of different proteins from its structure (induced by increased/decreased mechanical stress). The study subjects were fibers of the soleus muscle and cardiomyocytes of Wistar rats. Changes in external mechanical conditions were reconstructed by means of antiorthostatic suspension of the animals by their tails for 6, 12, 18, 24 and 72 hours. Transversal stiffness was measured by atomic force microscopy imaging; beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 levels in membranous and cytoplasmic fractions were quantified by Western blot analysis; expression rates of the corresponding genes were studied using RT-PCR. Results: In 6 hours, Alpha-Actinin 1 and Alpha-Actinin 4 levels decreased in the membranous fraction of proteins of cardiomyocytes and soleus muscle fibers, respectively, but increased in the cytoplasmic fraction of the abovementioned cells. After 6–12 hours of suspension, the expression rates of beta-, gamma-actin, Alpha-Actinin 1 and Alpha-Actinin 4 were elevated in the soleus muscle fibers, but the Alpha-Actinin 1 expression rate returned to the reference level in 72 hours. After 18–24 hours, the expression rates of beta-actin and Alpha-Actinin 4 increased in cardiomyocytes, while the Alpha-Actinin 1 expression rate decreased in soleus muscle fibers. After 12 hours, the beta- and gamma-actin content dropped in the membranous fraction and increased in the cytoplasmic protein fractions from both cardiomyocytes and soleus muscle fibers. The stiffness of both cell types decreased after the same period of time. Further, during the unloading period the concentration of nonmuscle actin and different isoforms of Alpha-Actinins increased in the membranous fraction from cardiomyocytes. At the same time, the concentration of the abovementioned proteins decreased in the soleus muscle fibers.
-
Structure and functional characteristics of rat's left ventricle cardiomyocytes under antiorthostatic suspension of various duration and subsequent reloading.
Journal of biomedicine & biotechnology, 2012Co-Authors: Irina V. Ogneva, Nikolay S. Biryukov, T. M. Mirzoev, O. M. Veselova, Irina M. LarinaAbstract:The goal of the research was to identify the structural and functional characteristics of the rat's left ventricle under antiorthostatic suspension within 1, 3, 7 and 14 days, and subsequent 3 and 7-day reloading after a 14-day suspension. The transversal stiffness of the cardiomyocyte has been determined by the atomic force microscopy, cell respiration—by polarography and proteins content—by Western blotting. Stiffness of the cortical cytoskeleton increases as soon as one day after the suspension and increases up to the 14th day, and starts decreasing during reloading, reaching the control level after 7 days. The stiffness of the contractile apparatus and the intensity of cell respiration also increases. The content of non-muscle isoforms of actin in the cytoplasmic fraction of proteins does not change during the whole experiment, as does not the beta-actin content in the membrane fraction. The content of gamma-actin in the membrane fraction correlates with the change in the transversal stiffness of the cortical cytoskeleton. Increased content of Alpha-Actinin-1 and Alpha-Actinin-4 in the membrane fraction of proteins during the suspension is consistent with increased gamma-actin content there. The opposite direction of change of Alpha-Actinin-1 and Alpha-Actinin-4 content suggests their involvement into the signal pathways.
Michael R. Ladomery - One of the best experts on this subject based on the ideXlab platform.
-
Kinetic behaviour of WT 1's zinc finger domain in binding to the Alpha-Actinin-1 mRNA.
Archives of Biochemistry and Biophysics, 2010Co-Authors: Elmar Nurmemmedov, Raymond K. Yengo, Michael R. Ladomery, Marjolein M. G. M. ThunnissenAbstract:The zinc finger transcription factor Wilms tumour protein (WT 1) is known for its essential involvement in the development of the genitourinary system as well as of other organs and tissues. WT 1 is capable of selectively binding either DNA or mRNA targets. A KTS insertion due to alternative splicing between the zinc fingers 3 and 4 and an unconventional zinc finger 1 are the unique features that distinguish WT 1 from classical DNA-binding C(2)H(2)-type zinc finger proteins. The DNA binding characteristics of WT 1 are well studied. Due to lack of information about its native RNA targets, no extensive research has been directed at how WT 1 binds RNA. Using surface plasmon resonance, this study attempts to understand the binding behaviour of WT 1 zinc fingers with its recently reported and first putative mRNA target, ACT 34, whose stem-loop structure is believed to be critical for the interactions with WT 1. We have analysed the interactions of five WT 1 zinc finger truncations with wild-type ACT 34 and four variants. Our results indicate that WT 1 zinc fingers bind ACT 34 in a specific manner, and that this occurs as interplay of all four zinc fingers. We also report that a sensitive kinetic balance, which is equilibrated by both zinc finger 1 and KTS, regulates the interaction with ACT 34. The stem-loop and the flanking nucleotides are important elements for specific recognition by WT 1 zinc fingers.
-
the wilms tumour suppressor protein wt1 kts isoform binds Alpha Actinin 1 mrna via its zinc finger domain
Biochemistry and Cell Biology, 2006Co-Authors: J P Venables, A A Morrison, Graham Dellaire, Michael R. LadomeryAbstract:Mutations in WT1 are associated with developmental syndromes that affect the urogenital system and neoplasms, including Wilms tumour, acute myeloid leukemia, and breast and prostate cancers. The WT1 protein belongs to the early growth response family of zinc-finger transcription factors. Uniquely to WT1, an evolutionarily conserved alternative splice event inserts the tripeptide KTS, between zinc fingers 3 and 4. Whereas –KTS isoforms bind DNA and activate or repress transcription, +KTS isoforms bind DNA less efficiently and interact with splice factors and RNA in vitro and in vivo. Although candidate DNA targets have been found, physiological mRNA targets are yet to be defined. We examined the distribution of WT1 in ribonucleoprotein (RNP) complexes in nuclear extract prepared from M15 cells, a mouse mesonephric fetal kidney cell line. WT1 cofractionated with the splice factor PSF in large RNP particles ≥2 MDa. We also found that PSF co-immunoprecipitated with WT1, suggesting a functional interaction bet...
A A Morrison - One of the best experts on this subject based on the ideXlab platform.
-
the wilms tumour suppressor protein wt1 kts isoform binds Alpha Actinin 1 mrna via its zinc finger domain
Biochemistry and Cell Biology, 2006Co-Authors: J P Venables, A A Morrison, Graham Dellaire, Michael R. LadomeryAbstract:Mutations in WT1 are associated with developmental syndromes that affect the urogenital system and neoplasms, including Wilms tumour, acute myeloid leukemia, and breast and prostate cancers. The WT1 protein belongs to the early growth response family of zinc-finger transcription factors. Uniquely to WT1, an evolutionarily conserved alternative splice event inserts the tripeptide KTS, between zinc fingers 3 and 4. Whereas –KTS isoforms bind DNA and activate or repress transcription, +KTS isoforms bind DNA less efficiently and interact with splice factors and RNA in vitro and in vivo. Although candidate DNA targets have been found, physiological mRNA targets are yet to be defined. We examined the distribution of WT1 in ribonucleoprotein (RNP) complexes in nuclear extract prepared from M15 cells, a mouse mesonephric fetal kidney cell line. WT1 cofractionated with the splice factor PSF in large RNP particles ≥2 MDa. We also found that PSF co-immunoprecipitated with WT1, suggesting a functional interaction bet...