scholarly journals Inhibition of the DNA-binding and transcriptional repression activity of the Wilms' tumor gene product, WT1, by cAMP-dependent protein kinase-mediated phosphorylation of Ser-365 and Ser-393 in the zinc finger domain

Oncogene ◽  
1997 ◽  
Vol 15 (17) ◽  
pp. 2001-2012 ◽  
Author(s):  
Yoshimasa Sakamoto ◽  
Mitsuaki Yoshida ◽  
Kentaro Semba ◽  
Tony Hunter
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Gene Product ◽  
Zinc Finger ◽  
Transcriptional Repression ◽  
Wilms Tumor ◽  
Zinc Finger Domain ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Tumor Gene

scholarly journals Overexpression of Zinc Finger Transcription Factor ZAT6 Enhances Salt Tolerance

2018 ◽  
Vol 13 (1) ◽  
pp. 431-445 ◽  
Author(s):  
Wei Tang ◽  
Caroline Luo
Keyword(s):  
Transcription Factor ◽  
Salt Stress ◽  
Protein Kinase ◽  
Stress Tolerance ◽  
Zinc Finger ◽  
Salt Stress Tolerance ◽  
Dependent Protein Kinase ◽  
Zinc Finger Transcription Factor ◽  
Transgenic Cell ◽  
Dependent Protein

AbstractThe purpose of the present investigation is to examine the function of the C2H2-type zinc finger transcription factor of Arabidopsis thaliana 6 (ZAT6) in salt stress tolerance in cells of rice (Oryza sativa L.), cotton (Gossypium hirsutum L.) and slash pine (Pinus elliottii Engelm.). Cells of O. sativa, G. hirsutum, and P. elliottii overexpressing ZAT6 were generated using Agrobacterium-mediated genetic transformation. Molecular and functional analysis of transgenic cell lines demonstrate that overexpression of ZAT6 increased tolerance to salt stress by decreasing lipid peroxidation and increasing the content of abscisic acid (ABA) and GA8, as well as enhancing the activities of antioxidant enzymes such as ascorbate peroxidise (APOX), catalase (CAT), glutathione reductase (GR), and superoxide dismutase (SOD). In rice cells, ZAT6 also increased expression of Ca2+-dependent protein kinase genes OsCPK9 and OsCPK25 by 5–7 fold under NaCl stress. Altogether, our results suggest that overexpression of ZAT6 enhanced salt stress tolerance by increasing antioxidant enzyme activity, hormone content and expression of Ca2+-dependent protein kinase in transgenic cell lines of different plant species.

Transcriptional repression mediated by the WT1 Wilms tumor gene product

Science ◽  
1991 ◽  
Vol 253 (5027) ◽  
pp. 1550-1553 ◽  
Author(s):  
S. Madden ◽  
D. Cook ◽  
J. Morris ◽  
A Gashler ◽  
V. Sukhatme ◽  
...  
Keyword(s):  
Gene Product ◽  
Transcriptional Repression ◽  
Wilms Tumor ◽  
Wilms Tumor Gene ◽  
Tumor Gene

scholarly journals Phosphorylation of SOX9 by Cyclic AMP-Dependent Protein Kinase A Enhances SOX9's Ability To Transactivate aCol2a1 Chondrocyte-Specific Enhancer

2000 ◽  
Vol 20 (11) ◽  
pp. 4149-4158 ◽  
Author(s):  
Wendong Huang ◽  
Xin Zhou ◽  
Véronique Lefebvre ◽  
Benoit de Crombrugghe
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Cyclic Amp ◽  
Protein Kinase A ◽  
Chondrocyte Differentiation ◽  
Phosphorylation Sites ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Kinase A

ABSTRACT Sox9 is a high-mobility-group domain-containing transcription factor required for chondrocyte differentiation and cartilage formation. We used a yeast two-hybrid method based on Son of Sevenless (SOS) recruitment to screen a chondrocyte cDNA library and found that the catalytic subunit of cyclic AMP (cAMP)-dependent protein kinase A (PKA-Cα) interacted specifically with SOX9. Next we found that two consensus PKA phosphorylation sites within SOX9 could be phosphorylated by PKA in vitro and that SOX9 could be phosphorylated by PKA-Cα in vivo. In COS-7 cells cotransfected with PKA-Cα and SOX9 expression plasmids, PKA enhanced the phosphorylation of wild-type SOX9 but did not affect phosphorylation of a SOX9 protein in which the two PKA phosphorylation sites (S64 and S211) were mutated. Using a phosphospecific antibody that specifically recognized SOX9 phosphorylated at serine 211, one of the two PKA phosphorylation sites, we demonstrated that addition of cAMP to chondrocytes strongly increased the phosphorylation of endogenous Sox9. In addition, immunohistochemistry of mouse embryo hind legs showed that Sox9 phosphorylated at serine 211 was principally localized in the prehypertrophic zone of the growth plate, corresponding to the major site of expression of the parathyroid hormone-related peptide (PTHrP) receptor. Since cAMP has previously been shown to effectively increase the mRNA levels of Col2a1 and other specific markers of chondrocyte differentiation in culture, we then asked whether PKA phosphorylation could modulate the activity of SOX9. Addition of 8-bromo-cAMP to chondrocytes in culture increased the activity of a transiently transfected SOX9-dependent 48-bp Col2a1chondrocyte-specific enhancer; similarly, cotransfection of PKA-Cα increased the activity of this enhancer. Mutations of the two PKA phosphorylation consensus sites of SOX9 markedly decreased the PKA-Cα activation of this enhancer by SOX9. PKA phosphorylation and the mutations in the consensus PKA phosphorylation sites of SOX9 did not alter its nuclear localization. In vitro phosphorylation of SOX9 by PKA resulted in more efficient DNA binding. We conclude that SOX9 is a target of cAMP signaling and that phosphorylation of SOX9 by PKA enhances its transcriptional and DNA-binding activity. Because PTHrP signaling is mediated by cAMP, our results support the hypothesis that Sox9 is a target of PTHrP signaling in the growth plate and that the increased activity of Sox9 might mediate the effect of PTHrP in maintaining the cells as nonhypertrophic chondrocytes.

scholarly journals Single-stranded-DNA binding alters human replication protein A structure and facilitates interaction with DNA-dependent protein kinase.

1996 ◽  
Vol 16 (9) ◽  
pp. 4798-4807 ◽  
Author(s):  
L J Blackwell ◽  
J A Borowiec ◽  
I A Mastrangelo
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Physical Interaction ◽  
Binding Modes ◽  
Dependent Protein Kinase ◽  
Single Stranded Dna ◽  
Dependent Protein

Human replication protein A (hRPA) is an essential single-stranded-DNA-binding protein that stimulates the activities of multiple DNA replication and repair proteins through physical interaction. To understand DNA binding and its role in hRPA heterologous interaction, we examined the physical structure of hRPA complexes with single-stranded DNA (ssDNA) by scanning transmission electron microscopy. Recent biochemical studies have shown that hRPA combines with ssDNA in at least two binding modes: by interacting with 8 to 10 nucleotides (hRPA8nt) and with 30 nucleotides (hRPA30nt). We find the relatively unstable hRPA8nt complex to be notably compact with many contacts between hRPA molecules. In contrast, on similar lengths of ssDNA, hRPA30nt complexes align along the DNA and make few intermolecular contacts. Surprisingly, the elongated hRPA30nt complex exists in either a contracted or an extended form that depends on ssDNA length. Therefore, homologous-protein interaction and available ssDNA length both contribute to the physical changes that occur in hRPA when it binds ssDNA. We used activated DNA-dependent protein kinase as a biochemical probe to detect alterations in conformation and demonstrated that formation of the extended hRPA30nt complex correlates with increased phosphorylation of the hRPA 29-kDa subunit. Our results indicate that hRPA binds ssDNA in a multistep pathway, inducing new hRPA alignments and conformations that can modulate the functional interaction of other factors with hRPA.

CsA-sensitive purine-box transcriptional regulator in bronchial epithelial cells contains NF45, NF90, and Ku

1998 ◽  
Vol 275 (6) ◽  
pp. L1164-L1172 ◽  
Author(s):  
Yosuke Aoki ◽  
Guohua Zhao ◽  
Daoming Qiu ◽  
Lingfang Shi ◽  
Peter N. Kao
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Dna Sequence ◽  
Transcriptional Activation ◽  
Nuclear Dna ◽  
Transcriptional Regulator ◽  
Catalytic Subunit ◽  
Dependent Protein Kinase ◽  
Bronchial Epithelial ◽  
Dependent Protein

Human bronchial epithelial (HBE) cells express interleukin (IL)-2 [Y. Aoki, D. Qiu, A. Uyei, and P. N. Kao. Am. J. Physiol. 272 ( Lung Cell. Mol. Physiol. 16): L276–L286, 1997]. 16HBE-transformed cells contain constitutive and inducible nuclear DNA-binding activity for the purine-box/nuclear factor (NF) of activated T cell (NFAT) target DNA sequence in the human IL-2 enhancer. Transcriptional activation through the purine-box DNA sequence requires stimulation with phorbol 12-myristate 13-acetate + ionomycin, and this activation is inhibited by cyclosporin A. Immunohistochemical staining of 16HBE cells demonstrates nuclear expression of the purine-box DNA-binding proteins NF45 and NF90 and no expression of NFATp or NFATc. NF90 and NF45 associate with the DNA-dependent protein kinase catalytic subunit and the DNA-targeting subunits Ku80 and Ku70 (N. S. Ting, P. N. Kao, D. W. Chan, L. G. Lintott, and S. P. Lees-Miller. J. Biol. Chem. 273: 2136–2145, 1998). Antibodies to Ku potently inhibit the purine-box DNA-binding complex. The purine-box transcriptional regulator in 16HBE cells likely comprises NF45, NF90, Ku80, Ku70, and the DNA-dependent protein kinase catalytic subunit.

scholarly journals Characterization of a Ku86 Variant Protein That Results in Altered DNA Binding and Diminished DNA-dependent Protein Kinase Activity

1996 ◽  
Vol 271 (24) ◽  
pp. 14098-14104 ◽  
Author(s):  
Zhiyong Han ◽  
Christine Johnston ◽  
Westley H. Reeves ◽  
Timothy Carter ◽  
James H. Wyche ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Kinase Activity ◽  
Protein Kinase Activity ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Variant Protein

scholarly journals The sak1+ gene of Schizosaccharomyces pombe encodes an RFX family DNA-binding protein that positively regulates cyclic AMP-dependent protein kinase-mediated exit from the mitotic cell cycle.

1995 ◽  
Vol 15 (3) ◽  
pp. 1479-1488 ◽  
Author(s):  
S Y Wu ◽  
M McLeod
Keyword(s):  
Protein Kinase ◽  
Dna Binding ◽  
Cyclic Amp ◽  
Stationary Phase ◽  
Schizosaccharomyces Pombe ◽  
Sexual Differentiation ◽  
Loss Of Function ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Diploid Cells

In Schizosaccharomyces pombe, meiosis is initiated by conditions of nutrient deprivation. Mutations in genes encoding elements of the cyclic AMP-dependent protein kinase (cAPK) pathway interfere with meiosis. Loss-of-function alleles of genes that stimulate the activity of cAPK allow cells to bypass the normal requirement of starvation for conjugation and meiosis. Alternatively, loss-of-function alleles of genes that inhibit cAPK lead to the inability to undergo sexual differentiation. The cgs1+ gene encodes the regulatory subunit of cAPK, and the cgs2+ gene encodes a cyclic AMP phosphodiesterase. Thus, both genes encode proteins which negatively regulate the activity of cAPK. Loss of either cgs1 or cgs2 prevents haploid cells from conjugating and diploid cells from undergoing meiosis. In addition to these defects, cells are unable to enter stationary phase. We describe a novel gene, sak1+, which when present on a plasmid overcomes the aberrant phenotypes associated with unregulated cAPK activity. Genetic analysis of sak1+ (suppressor of A-kinase) reveals that it functions downstream of cyclic AMP-dependent protein kinase to allow cells to exist the mitotic cycle and enter either stationary phase or the pathway leading to sexual differentiation. The sak1+ gene is essential for cell viability, and a null allele causes multiple defects in cell morphology and nuclear division. Thus, sak1+ is an important regulatory element in the life cycle of S. pombe. Sequence analysis shows that the predicted product of the sak1+ gene is an 87-kDa protein which shares homology to the RFX family of DNA-binding proteins identified in humans and mice. One member of this family, RFX1, is a transcription factor for a variety of viral and cellular genes.

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