scholarly journals CAD Protein

2020 ◽  
Author(s):  
Keyword(s):  
Cad Protein

Transcriptional regulation of the human CAD gene during myeloid differentiation

1987 ◽  
Vol 7 (5) ◽  
pp. 1961-1966
Author(s):  
G N Rao ◽  
E S Buford ◽  
J N Davidson
Keyword(s):  
Gene Expression ◽  
De Novo ◽  
Morphological Changes ◽  
Transcriptional Level ◽  
Carbamoyl Phosphate ◽  
Aspartate Transcarbamylase ◽  
Trans Retinoic Acid ◽  
Beta Actin ◽  
Cad Gene ◽  
Cad Protein

CAD codes for a trifunctional protein involved in the catalysis of the first three enzymatic activities in the de novo pyrimidine biosynthetic pathway, namely, carbamoyl-phosphate synthetase II (EC 6.3.5.5), aspartate transcarbamylase (EC 2.1.3.2), and dihydroorotase (EC 3.5.2.3). CAD regulation was studied in the human promyelocyte leukemic line HL-60 as it differentiated into monocytic or granulocytic lineages after induction by 12-O-tetradecanoylphorbol-13-acetate or trans-retinoic acid and dibutyryl cyclic AMP, respectively. Within 12 h of induction of HL-60 cells with either inducer, total cellular levels of CAD RNA essentially disappeared. On the other hand, no apparent decreases in beta-actin RNA levels were seen even 48 h after HL-60 cells were induced, as compared with untreated cells. With nuclear runoff assays, it was clearly shown that the inactivation of CAD gene expression during the induction of HL-60 cells with either inducer was at the transcriptional level. The nuclear runoff experiments also demonstrated that the CAD gene expression was shut down in less than 4 h after induction, well before morphological changes were observed in these cells. At the enzymatic level, the activity of aspartate transcarbamylase, one of the three enzymes encoded by the CAD gene, decreased by about half within 24 h of induction, suggesting a CAD protein half-life of 24 h in differentiating HL-60 cells. Nevertheless, this means that significant levels of aspartate transcarbamylase activity remained even after the cells have stopped proliferating. From the RNA data, it is clear that CAD gene expression is rapidly turned off as promyelocytes begin to terminally differentiate into macrophages and granulocytes. We suspect that the inactivation of the CAD gene in induced HL-60 cells is a consequence of the differentiating cells leaving the cell cycle and becoming nonproliferating.

Effects of ectopic expression of caudal during Drosophila development

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 271-277 ◽  
Author(s):  
M. Mlodzik ◽  
G. Gibson ◽  
W.J. Gehring
Keyword(s):  
Heat Shock ◽  
Imaginal Disc ◽  
Ectopic Expression ◽  
Homeobox Gene ◽  
Homeotic Genes ◽  
Posterior Half ◽  
Fushi Tarazu ◽  
Drosophila Development ◽  
Head Development ◽  
Cad Protein

The effects of heat-shock-induced ectopic expression of the homeobox gene caudal (cad) at all stages of Drosophila development have been examined. Presence of cad protein (CAD) at the anterior end of cellular blastoderm embryos was found to disrupt head development and segmentation, due to alteration of the expression of segmentation genes such as fushi tarazu and engrailed, as well as repression of head-determining genes such as Deformed. These results support the conclusion that, while CAD is probably required to activate transcription of fushi tarazu in the posterior half of the embryo, it should not be expressed in the anterior half prior to gastrulation, and thus suggest a role for the CAD gradient. Ectopic expression of CAD at later stages of development has no obvious effects on embryogenesis or imaginal disc development, suggesting that the homeotic genes of the Antennapedia and Bithorax Complexes are almost completely epistatic to caudal.

Construction of a cDNA to the hamster CAD gene and its application toward defining the domain for aspartate transcarbamylase

1985 ◽  
Vol 5 (7) ◽  
pp. 1735-1742
Author(s):  
K Shigesada ◽  
G R Stark ◽  
J A Maley ◽  
L A Niswander ◽  
J N Davidson
Keyword(s):  
Protein A ◽  
Aspartate Transcarbamylase ◽  
Native Form ◽  
Multifunctional Protein ◽  
E Coli ◽  
Bacterial Enzyme ◽  
Cad Gene ◽  
Mammalian Enzyme ◽  
Quaternary Structures ◽  
Cad Protein

cDNA complementary to hamster mRNA encoding the CAD protein, a multifunctional protein which carries the first three enzymes of pyrimidine biosynthesis, was constructed. The longest of these recombinants (pCAD142) covers 82% of the 7.9-kilobase mRNA. Portions of the cDNA were excised and replaced by a lac promoter-operator-initiation codon segment. The resultant plasmids were transfected into an Escherichia coli mutant defective in aspartate transcarbamylase, the second enzyme of the pathway. Complementation of the bacterial defect was observed with as little as 2.2 kilobases of cDNA sequence, corresponding to the 3' region of the mRNA. DNA sequencing in this region of the hamster cDNA reveals stretches which are highly homologous to the E. coli gene for the catalytic subunit of aspartate transcarbamylase; other stretches show no homology. The highly conserved regions probably reflect areas of protein structure critical to catalysis, while the nonconserved regions may reflect differences between the quaternary structures of E. coli and mammalian aspartate transcarbamylases, one such difference being that the bacterial enzyme in its native form is allosterically regulated and the mammalian enzyme is not.

Synthesis of the nonconserved dihydroorotase domain of the multifunctional hamster CAD protein in Escherichia coli

Gene ◽  
1991 ◽  
Vol 99 (2) ◽  
pp. 211-216 ◽  
Author(s):  
Lisa A. Musmanno ◽  
Julie A. Maley ◽  
Jeffrey N. Davidson
Keyword(s):  
Escherichia Coli ◽  
Cad Protein

scholarly journals Substitutions in hamster CAD carbamoyl-phosphate synthetase alter allosteric response to 5-phosphoribosyl-alpha-pyrophosphate (PRPP) and UTP

2004 ◽  
Vol 378 (3) ◽  
pp. 991-998 ◽  
Author(s):  
Christine Q. SIMMONS ◽  
Alan J. SIMMONS ◽  
Aaron HAUBNER ◽  
Amber REAM ◽  
Jeffrey N. DAVIDSON
Keyword(s):  
De Novo ◽  
Binding Pocket ◽  
Tryptophan Residue ◽  
Carbamoyl Phosphate Synthetase ◽  
Carbamoyl Phosphate ◽  
Aspartate Transcarbamylase ◽  
Inosine Monophosphate ◽  
E Coli ◽  
Mammalian Enzyme ◽  
Cad Protein

CPSase (carbamoyl-phosphate synthetase II), a component of CAD protein (multienzymic protein with CPSase, aspartate transcarbamylase and dihydro-orotase activities), catalyses the regulated steps in the de novo synthesis of pyrimidines. Unlike the orthologous Escherichia coli enzyme that is regulated by UMP, inosine monophosphate and ornithine, the mammalian CPSase is allosterically inhibited by UTP, and activated by PRPP (5-phosphoribosyl-α-pyrophosphate) and phosphorylation. Four residues (Thr974, Lys993, Lys954 and Thr977) are critical to the E. coli inosine monophosphate/UMP-binding pocket. In the present study, three of the corresponding residues in the hamster CPSase were altered to determine if they affect either PRPP activation or UTP inhibition. Substitution of the hamster residue, positionally equivalent to Thr974 in the E. coli enzyme, with alanine residue led to an enzyme with 5-fold lower activity and a near loss of PRPP activation. Whereas replacement of the tryptophan residue at position 993 had no effect, an Asp992→Asn substitution yielded a much-activated enzyme that behaved as if PRPP was present. The substitution Lys954→Glu had no effect on PRPP stimulation. Only modest decreases in UTP inhibitions were observed with each of the altered CPSases. The results also show that while PRPP and UTP can act simultaneously, PRPP activation is dominant. Apparently, UTP and PRPP have distinctly different associations within the mammalian enzyme. The findings of the present study may prove relevant to the neuropathology of Lesch–Nyhan syndrome.

scholarly journals Transcriptional regulation of the human CAD gene during myeloid differentiation.

1987 ◽  
Vol 7 (5) ◽  
pp. 1961-1966 ◽  
Author(s):  
G N Rao ◽  
E S Buford ◽  
J N Davidson
Keyword(s):  
Gene Expression ◽  
De Novo ◽  
Morphological Changes ◽  
Transcriptional Level ◽  
Carbamoyl Phosphate ◽  
Aspartate Transcarbamylase ◽  
Trans Retinoic Acid ◽  
Beta Actin ◽  
Cad Gene ◽  
Cad Protein

CAD codes for a trifunctional protein involved in the catalysis of the first three enzymatic activities in the de novo pyrimidine biosynthetic pathway, namely, carbamoyl-phosphate synthetase II (EC 6.3.5.5), aspartate transcarbamylase (EC 2.1.3.2), and dihydroorotase (EC 3.5.2.3). CAD regulation was studied in the human promyelocyte leukemic line HL-60 as it differentiated into monocytic or granulocytic lineages after induction by 12-O-tetradecanoylphorbol-13-acetate or trans-retinoic acid and dibutyryl cyclic AMP, respectively. Within 12 h of induction of HL-60 cells with either inducer, total cellular levels of CAD RNA essentially disappeared. On the other hand, no apparent decreases in beta-actin RNA levels were seen even 48 h after HL-60 cells were induced, as compared with untreated cells. With nuclear runoff assays, it was clearly shown that the inactivation of CAD gene expression during the induction of HL-60 cells with either inducer was at the transcriptional level. The nuclear runoff experiments also demonstrated that the CAD gene expression was shut down in less than 4 h after induction, well before morphological changes were observed in these cells. At the enzymatic level, the activity of aspartate transcarbamylase, one of the three enzymes encoded by the CAD gene, decreased by about half within 24 h of induction, suggesting a CAD protein half-life of 24 h in differentiating HL-60 cells. Nevertheless, this means that significant levels of aspartate transcarbamylase activity remained even after the cells have stopped proliferating. From the RNA data, it is clear that CAD gene expression is rapidly turned off as promyelocytes begin to terminally differentiate into macrophages and granulocytes. We suspect that the inactivation of the CAD gene in induced HL-60 cells is a consequence of the differentiating cells leaving the cell cycle and becoming nonproliferating.

scholarly journals Construction of a cDNA to the hamster CAD gene and its application toward defining the domain for aspartate transcarbamylase.

1985 ◽  
Vol 5 (7) ◽  
pp. 1735-1742 ◽  
Author(s):  
K Shigesada ◽  
G R Stark ◽  
J A Maley ◽  
L A Niswander ◽  
J N Davidson
Keyword(s):  
Protein A ◽  
Aspartate Transcarbamylase ◽  
Native Form ◽  
Multifunctional Protein ◽  
E Coli ◽  
Bacterial Enzyme ◽  
Cad Gene ◽  
Mammalian Enzyme ◽  
Quaternary Structures ◽  
Cad Protein

cDNA complementary to hamster mRNA encoding the CAD protein, a multifunctional protein which carries the first three enzymes of pyrimidine biosynthesis, was constructed. The longest of these recombinants (pCAD142) covers 82% of the 7.9-kilobase mRNA. Portions of the cDNA were excised and replaced by a lac promoter-operator-initiation codon segment. The resultant plasmids were transfected into an Escherichia coli mutant defective in aspartate transcarbamylase, the second enzyme of the pathway. Complementation of the bacterial defect was observed with as little as 2.2 kilobases of cDNA sequence, corresponding to the 3' region of the mRNA. DNA sequencing in this region of the hamster cDNA reveals stretches which are highly homologous to the E. coli gene for the catalytic subunit of aspartate transcarbamylase; other stretches show no homology. The highly conserved regions probably reflect areas of protein structure critical to catalysis, while the nonconserved regions may reflect differences between the quaternary structures of E. coli and mammalian aspartate transcarbamylases, one such difference being that the bacterial enzyme in its native form is allosterically regulated and the mammalian enzyme is not.

scholarly journals The Effects of Rhubarb for the Treatment of Diabetic Nephropathy in Animals: A Systematic Review and Meta-analysis

2021 ◽  
Vol 12 ◽  
Author(s):  
Jing-Yi Zeng ◽  
Yu Wang ◽  
Miao Miao ◽  
Xiao-Rong Bao
Keyword(s):  
Systematic Review ◽  
Diabetic Nephropathy ◽  
Clinical Application ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Meta Analysis ◽  
Cochrane Library ◽  
Tubulointerstitial Injury ◽  
Alpha Smooth Muscle Actin ◽  
Cad Protein

Background: Rhubarb, also known as Da Huang, is a traditional Chinese medicine, and it was often used as a laxative in the past. Recently, multiple studies have applied rhubarb to treat diabetic nephropathy (DN). Anthraquinones, including emodin and rhein, have been extracted from rhubarb and used to explore the effective components and possible mechanisms of rhubarb for DN. Evaluating the efficacy of rhubarb may provide a scientific reference for the clinical application of rhubarb for the treatment of DN.Objective: 1) To evaluate the efficacy of rhubarb in the treatment of DN; 2) To identify the most effective ingredient of rhubarb for DN; 3) To explore the specific mechanism of rhubarb in treating DN.Methods: Data sources: related studies were identified by searching Cochrane Library, Ovid-EMBASE, PubMed, SinoMed, WanFang, VIP, CNKI, and other Chinese magazines.Assessment and analysis: SYRCLE’s risk of bias tool for animal studies was used to assess the quality of articles. The meta-analysis was performed in accordance with the Cochrane Handbook for Systematic Reviews of Interventions. Data analysis adopted RevMan 5.3 and STATA 12.0 software.This study was published in the register with PROSPERO, number CRD42020204701.Results: Aggregated data were collected from 27 eligible studies. The results illustrated an intense improvement in the following outcomes in rhubarb-treated animals with DN (p < 0.05): blood glucose, serum creatinine (Scr), blood urea nitrogen (BUN), albumin creatinine ratio (ACR), urine protein (UP), urinary albumin excretion (UAE), renal index (two kidneys weight/body weight, KW/BW), tubulointerstitial injury index (TII), transforming growth factor-beta1 (TGF-β1) mRNA and protein, alpha-smooth muscle actin (α-SMA) protein, and E-cadherin (E-cad) protein. Of these, DN animals with rhubarb exhibited a significantly higher level of E-cad protein. In addition, the level of the other outcomes mentioned above decreased significantly, while there was no significant association between the intervention and nephrin protein (p > 0.05).Conclusion: This systematic review and meta-analysis demonstrated that rhubarb has a positive therapeutic effect on animals with DN, which may provide confidence and some theoretical reference for clinical application to a certain extent.

ERK1/2-mediated phosphorylation of myometrial caldesmon during pregnancy and labor

2003 ◽  
Vol 284 (1) ◽  
pp. R192-R199 ◽  
Author(s):  
Yunping Li ◽  
Hyun-Dong Je ◽  
Sabah Malek ◽  
Kathleen G. Morgan
Keyword(s):  
Protein Content ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Fold Increase ◽  
Myosin Light Chain Phosphorylation ◽  
Force Curve ◽  
Pregnant Rat ◽  
Myometrial Contractility ◽  
Cad Protein ◽  
In Pregnancy

We used a timed-pregnant rat model to track changes in myometrial contractility during pregnancy and labor and to correlate these changes with upstream signaling events. Myometrium was harvested from CO2-euthanized rats. Although contraction amplitudes increased at 16 and 20 days of pregnancy, contraction incidence and area under the force curve were inhibited, consistent with the myometrial quiescence of pregnancy. The Ca2+ sensitivity of contraction was decreased at 20 days of pregnancy and this was partially reversed in labor. The protein content of h-caldesmon ( h-CaD) was increased in pregnancy. A 40-fold increase in the signal from a phospho-CaD antibody specific for phosphorylation at an ERK1/2 site occurred during labor. ERK1/2 activation increased significantly at the onset of labor. Myosin light chain phosphorylation (LC20-P) increased significantly in labor compared with the nonpregnant state. Thus we conclude that the increase in CaD protein content during pregnancy may contribute to a suppression of the contractility of pregnant myometrium. Conversely, CaD phosphorylation, through an ERK1/2-mediated signaling pathway, as well as an increase in basal LC20-P, is suggested to contribute to the reversal of inhibition and promote contraction of the uterus during labor.

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