Characterization of Transcriptional Regulatory Elements in the Promoter Region of the Murine Blood Coagulation Factor VII Gene

Stauffer, Daniel; Chukwumezie, Beatrice N.; Wilberding, Julie A.; Rosen, Elliot D.

doi:10.1074/jbc.273.4.2277

Cited by 29 publications

(27 citation statements)

References 80 publications

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“…The presence of the anti-HNF4␣ antibody resulted in a much lower binding of the receptor to the 1A9HNF4␣ REwt probe (lane 3) compared with the apo CIII probe (lane 6). However, lower exposure of the shift revealed a similar decrease of the binding onto the apo CIII probe in the presence of the antibody (data not shown), and a similar reduction of the binding of HNF4␣ to DNA probes in the presence of the antibody was reported in various studies (Stauffer et al, 1998;Stroup and Chiang, 2000;Wang et al, 2000;Bartoov-Shifman et al, 2002). However, no protein-DNA complex was observed when using the mutated HNF4␣ RE probe (Fig.…”

Section: Hnf4␣ Controls Ugt1a9 Expression 243supporting

confidence: 85%

Hepatic Expression of the UGT1A9 Gene Is Governed by Hepatocyte Nuclear Factor 4α

Barbier¹,

Girard²,

Inoue³

et al. 2004

Mol Pharmacol

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UDP-glucuronosyltransferase (UGT) enzymes catalyze the glucuronidation reaction, which is a major pathway in the catabolism and elimination of numerous endo-and xenobiotics. Among the UGT enzyme family members, the UGT1A7, UGT1A8, UGT1A9, and UGT1A10 isoforms are issued from a single gene through differential splicing. However, these enzymes display distinct tissue-specific expression patterns. Indeed, UGT1A7, UGT1A8, and UGT1A10 are exclusively expressed in extrahepatic tissues, whereas UGT1A9 transcripts are found at high concentrations in liver. In the present study, we report that the liver-enriched hepatocyte nuclear factor 4 (HNF4)-␣ controls the hepatic expression of the UGT1A9 enzyme. Liver-specific disruption of the HNF4␣ gene in mice drastically decreases liver UGT1A9 mRNA levels. Furthermore, an HNF4␣ response element (HNF4␣ RE) was identified in the promoter of human UGT1A9 at position Ϫ372 to Ϫ360 base pairs by transient transfection, electrophoretic mobility shift assays, and chromatin immunoprecipitation experiments. It is interesting that this response element is absent in the proximal UGT1A7, UGT1A8, and UGT1A10 gene promoters. In conclusion, the present study identifies HNF4␣ as a major factor for the control of UGT1A9 hepatic expression and suggests that the absence of UGT1A7, UGT1A8, and UGT1A10 expression in the liver is caused by, at least in part, a few base pair changes in their promoter sequences in the region corresponding to the HNF4␣ RE of the UGT1A9 gene.

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Section: Hnf4␣ Controls Ugt1a9 Expression 243supporting

confidence: 85%

Hepatic Expression of the UGT1A9 Gene Is Governed by Hepatocyte Nuclear Factor 4α

Barbier¹,

Girard²,

Inoue³

et al. 2004

Mol Pharmacol

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“…2). However, daily mean FVII expression levels in Clock Ϫ/Ϫ ; Npas2 Ϫ/Ϫ mice were not significantly different from those detected in wild-type mice (two-way ANOVA; F (1,50) ϭ 1.48, P Ͼ 0.2), highlighting the importance of the other cis/trans elements responsible for the constitutive expression of mouse FVII gene (39).…”

Section: Resultsmentioning

confidence: 73%

“…For these studies, we created a luciferase reporter construct that contains the 5Ј regulatory region of mouse FVII gene (1,087 bp) including the E-box elements (Fig. 5A) and that was previously shown to possess promoter activity (39). Because of the liver-specific expression of FVII, the mouse hepatoma cell line Hepa1-6 was used for reporter expression analysis.…”

Section: Transactivation Of Mouse Fvii Promoter By Circadian Clock Comentioning

confidence: 99%

Evidence for an Overlapping Role of CLOCK and NPAS2 Transcription Factors in Liver Circadian Oscillators

Bertolucci

Cavallari

Colognesi

et al. 2008

Molecular and Cellular Biology

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The mechanisms underlying the circadian control of gene expression in peripheral tissues and influencing many biological pathways are poorly defined. Factor VII (FVII), the protease triggering blood coagulation, represents a valuable model to address this issue in liver since its plasma levels oscillate in a circadian manner and its promoter contains E-boxes, which are putative DNA-binding sites for CLOCK-BMAL1 and NPAS2-BMAL1 heterodimers and hallmarks of circadian regulation. The peaks of FVII mRNA levels in livers of wild-type mice preceded those in plasma, indicating a transcriptional regulation, and were abolished in Clock ؊/؊ ; Npas2 ؊/؊ mice, thus demonstrating a role for CLOCK and NPAS2 circadian transcription factors. The investigation of Npas2؊/؊ and Clock ⌬19/⌬19 mice, which express functionally defective heterodimers, revealed robust rhythms of FVII expression in both animal models, suggesting a redundant role for NPAS2 and CLOCK. The molecular bases of these observations were established through reporter gene assays. FVII transactivation activities of the NPAS2-BMAL1 and CLOCK-BMAL1 heterodimers were (i) comparable (a fourfold increase), (ii) dampened by the negative circadian regulators PER2 and CRY1, and (iii) abolished upon E-box mutagenesis. Our data provide the first evidence in peripheral oscillators for an overlapping role of CLOCK and NPAS2 in the regulation of circadianly controlled genes.Circadian clocks are endogenous oscillators that drive biochemical, physiological, and behavioral processes in organisms (36), and the alteration of which is associated with human diseases (17,26,42). It is well documented that the circadian rhythms of mammals are controlled by a master circadian pacemaker located in the suprachiasmatic nucleus (SCN) of the anterior hypothalamus (38) as well as by peripheral oscillators located in most tissues (43).The basic circadian molecular clockworks consist of interacting positive and negative transcriptional/translational feedback loops (11). The positive loop involves CLOCK-BMAL1 and NPAS2-BMAL1 heterodimers, three basic helix-loop-helix transcriptional activators that bind to E-box elements (CA CGTG) located in the regulatory regions of the Period (Per1, Per2, and Per3) and Cryptochrome (Cry1 and Cry2) genes (3, 13, 16, 24). CRY and PER proteins form oligomers that, once transported into the nucleus, repress their own transcription by inhibiting CLOCK-BMAL1 or NPAS2-BMAL1 (25). Furthermore, other transcription factors (CIPC, DEC1, DEC2, and Fbxl3) play important roles in the negative-feedback loop (15,19,44). The positive and negative limbs are interlaced: CLOCK-BMAL1 heterodimers indirectly regulate a rhythm in Bmal1 transcription; the nuclear orphan receptor genes Reverb␣ and Rora are coordinately activated by CLOCK-BMAL1 to produce proteins that compete for the same promoter element but have opposing actions on Bmal1 transcription (1, 32). Oscillations in gene expression resulting from this feedback loop have period lengths of approximately 24 h, thus giving ri...

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“…Steady State Mg 2ϩ -ATPase Measurements-ATPase assays were performed essentially as described previously (15) in buffer A (20 mM Hepes, pH 7.4, 1 mM MgCl 2 , 2 mM EGTA, 27 mM KCl, and 1 mM ␤-mercaptoethanol) or buffer B (15 mM KCl, 1 mM EGTA, 1.5 mM MgCl 2 , 10 mM Hepes, pH 7.4, 150 mM imidazole, 25 mM NaCl, 5% glycerol, 10 mM Tris/Cl, pH 8.0, 1 mM ␤-mercaptoethanol) when Myo9b proteins were assayed after the elution from the HiTrap column directly.…”

Section: Myo9bmentioning

confidence: 99%

Kinetic Mechanism of Myosin IXB and the Contributions of Two Class IX-specific Regions

Nalavadi

Nyitrai

Bertolini

et al. 2005

Journal of Biological Chemistry

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Myosin IXb (Myo9b) was reported to be a single-headed, processive myosin. In its head domain it contains an N-terminal extension and a large loop 2 insertion that are specific for class IX myosins. We characterized the kinetic properties of purified, recombinant rat Myo9b, and we compared them with those of Myo9b mutants that had either the N-terminal extension or the loop 2 insertion deleted. Unlike other processive myosins, Myo9b exhibited a low affinity for ADP, and ADP release was not rate-limiting in the ATPase cycle. Myo9b is the first myosin for which ATP hydrolysis or an isomerization step after ATP binding is rate-limiting. Myo9b-ATP appeared to be in a conformation with a weak affinity for actin as determined by pyrene-actin fluorescence. However, in actin cosedimentation experiments, a subpopulation of Myo9b-ATP bound F-actin with a remarkably high affinity. Deletion of the N-terminal extension reduced actin affinity and increased the rate of nucleotide binding. Deletion of the loop 2 insertion reduced the actin affinity and altered the communication between actin and nucleotide-binding sites.The myosins represent a large superfamily of motor molecules that convert the chemical energy liberated by ATP hydrolysis into mechanical force along actin filaments. They have been subdivided into 18 different classes based on homologous myosin head domain sequences (1). In addition to a characteristic head domain, all myosins contain a light chain binding domain and a tail domain. The tail domains of some myosins dimerize giving rise to two-headed myosins. Motor properties like direction of movement, speed, step size, duty ratio, processivity, and regulation can vary greatly between various myosins. Movement on actin involves the repeated hydrolysis of ATP by myosins, leading to an ordered cycling between different nucleotide binding states that exhibit different actin binding affinities. Cycling rates and relative time spent in these nucleotide states vary in different myosins, such that the fraction of time a given myosin remains strongly attached to the actin filament during its ATPase cycle differs substantially. Generally, myosin-ATP and myosin-ADP-P i bind weakly to actin filaments, whereas myosin-ADP and myosin alone bind strongly (2). In many myosins, e.g. skeletal muscle myosin II, the release of P i is the rate-limiting step, which means that they spend a large fraction of the total cycling time in a weak actinbinding state. On the other hand, in class V and class VI myosins, the cycle is modified in a way that ADP release is slow and rate-limiting, so these myosins spend most of their cycling time strongly attached to actin (3, 4). Myosin V is two-headed and the two heads cooperate to allow for continuous, processive, hand over hand movement along actin filaments (5).The class IX myosin, myosin 9b (Myo9b), 5 has been reported to exhibit unique motor properties. Despite being a single-headed myosin, it demonstrated typical characteristics of a processive myosin in in vitro motility assays, meaning tha...

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Characterization of Transcriptional Regulatory Elements in the Promoter Region of the Murine Blood Coagulation Factor VII Gene

Cited by 29 publications

References 80 publications

Hepatic Expression of the UGT1A9 Gene Is Governed by Hepatocyte Nuclear Factor 4α

Hepatic Expression of the UGT1A9 Gene Is Governed by Hepatocyte Nuclear Factor 4α

Evidence for an Overlapping Role of CLOCK and NPAS2 Transcription Factors in Liver Circadian Oscillators

Kinetic Mechanism of Myosin IXB and the Contributions of Two Class IX-specific Regions

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