Transcription factor specificity protein 1 (Sp1) is the main regulator of nerve growth factor‐induced sphingosine kinase 1 gene expression of the rat pheochromocytoma cell line, PC12

Sobue, Satoshi; Hagiwara, Kaoru; Banno, Yoshiko; Tamiya‐Koizumi, Keiko; Suzuki, Motoshi; Takagi, Akira; Kojima, Tetsuhito; Asano, Haruhiko; Nozawa, Yoshinori; Murate, Takashi

doi:10.1111/j.1471-4159.2005.03399.x

Cited by 56 publications

(64 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The cells were cultured in 60-mm dishes until they became 50% confluent, and then were transiently transfected with 10 mg of the expression plasmid vectors using the calcium phosphate method as described previously [10]. After incubation in FCS-free DMEM for 24 hr, the cell culture media and cell lysates that had dissolved in reporter lysis buffer (Promega, Madison, USA) were collected, centrifuged at Â 1500g for 10 min, and prepared as Western blot samples.…”

Section: Transient Expression Of the Recombinant Atsmentioning

confidence: 99%

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

et al. 2007

View full text Add to dashboard Cite

We analyzed the antithrombin (AT) gene in four unrelated Japanese patients with an AT deficiency, and individually identified four distinct mutations in the heterozygous state. There were two novel mutations, 2417delT leading to a frameshift with a premature termination at amino acid -3 (FS-3Stop) and C2640T resulting in a missense mutation (Ala59Val). Previously reported mutations, T5342C (Ser116Pro) and T72C (Met-32Thr), were also found in the other two patients. To understand the molecular basis responsible for the AT deficiency in these patients, in vitro expression experiments were performed using HEK293 cells transfected with either wild type or respective mutant AT expression vector. We found that -3Stop-AT and -32Thr-AT were not secreted into the culture media, whereas 116Pro-AT and 59Val-AT were secreted normally. We further studied the heparin cofactor activity and the binding to heparin of each recombinant AT molecule. Ser116Pro mutation significantly impaired the binding affinity to heparin resulting in a reduced heparin cofactor activity. In contrast, we found that Ala59Val mutant AT unexpectedly showed a normal affinity to heparin, but severely impaired the heparin cofactor activity. Our findings suggested that FS-3Stop and Met-32Thr mutations are responsible for type I AT deficiency, whereas Ser116Pro and Ala59Val mutations contribute to type II AT deficiency, confirming that there were diverse molecular mechanisms of AT deficiency depend upon discrete AT gene abnormalities as reported previously. Am. J. Hematol. 82:702-705, 2007. V

show abstract

Section: Transient Expression Of the Recombinant Atsmentioning

confidence: 99%

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

et al. 2007

View full text Add to dashboard Cite

show abstract

“…DNA transfection, establishment of stable transfectants and Western blotting DNA transfection was performed by a calcium precipitation method as described previously (Sobue et al, 2005). Stable transfectants of SH-SY5Y cell were established by G418 selection.…”

Section: Methodsmentioning

confidence: 99%

“…EMSA was performed according to the method described previously (Sobue et al, 2005). For the supershift experiment, anti-Fli-1 (Santa Cruz Inc.) or anti-EWS antibody (Santa Cruz Inc.) was mixed with nuclear extract for 15 min at room temperature before mixing with the labeled probe as described below.…”

Section: Semi-quantitative Rt-pcrmentioning

confidence: 99%

See 1 more Smart Citation

Ewing's sarcoma fusion protein, EWS/Fli-1 and Fli-1 protein induce PLD2 but not PLD1 gene expression by binding to an ETS domain of 5′ promoter

et al. 2006

View full text Add to dashboard Cite

It was reported that short interfering RNA (siRNA) of EWS/Fli-1 downregulated phospholipase D (PLD)2 in Ewing's sarcoma (EWS) cell line, suggesting that PLD2 is the target of aberrant transcription factor, EWS/Fli-1. Here, we further investigated the regulation of PLD2 gene expression by EWS/Fli-1 and Fli-1 in another EWS cell line, and also in EWS/Fli-1-or Fli-1-transfected cell line. EWS/Fli-1-or Fli-1-overexpressed cells showed higher PLD2 but not PLD1 protein expression and enhanced cell proliferation as compared to mock transfectant. The treatment of these cells with 1-butanol or siRNA of PLD2 inhibited cell growth, suggesting the pivotal role of PLD in cell growth promotion. PLD2 but not PLD1 mRNA level was also increased in EWS/Fli-1 or Fli-1-transfectants. After determining the transcription initiation points, we cloned the 5 0 promoter of both PLD1 and PLD2 and analysed promoter activities. Results showed that EWS/ Fli-1 and Fli-1 increase PLD2 gene expression by binding to an erythroblast transformation-specific domain (À126 to À120 bp from the transcription initiation site) of PLD2 promoter, which is the minimal and most powerful region. Electrophoresis mobility shift assay using truncated proteins showed that both DNA-binding domain and trans-activating domain were necessary for the enhanced gene expression of PLD2.

show abstract

“…It contains a mannose-specific carbohydrate recognition domain (CRD) on the ER luminal side, and ER exit and retrieval motifs on the cytoplasmic side [8]. MCFD2 has an EF-hand domain that interacts with LMAN1 in a Ca 21 -dependent manner [5]. The LMAN1-MCFD2 protein complex functions as a cargo receptor that facilitates the transport of FV and FVIII from the ER to the Golgi apparatus [5,9].…”

Section: Introductionmentioning

confidence: 99%

A novel missense mutation causing abnormal LMAN1 in a Japanese patient with combined deficiency of factor V and factor VIII

et al. 2009

View full text Add to dashboard Cite

Combined deficiency of coagulation factor V (FV) and factor VIII (FVIII) (F5F8D) is an inherited bleeding disorder characterized by a reduction in plasma concentrations of FV and FVIII. F5F8D is genetically linked to mutations in either LMAN1 or MCFD2. Here, we investigated the molecular basis of F5F8D in a Japanese patient, and identified a novel missense mutation (p.Trp67Ser, c.200G>C) in the LMAN1, but no mutation in the MCFD2. The amount of LMAN1 in Epstein-Barr virus-immortalized lymphoblasts from the patient was found to be almost the same as that in cells from a normal individual. Interestingly, an anti-MCFD2 antibody did not co-immunoprecipitate the mutant LMAN1 with MCFD2 in lymphoblasts from the patient, suggesting the affinity of MCFD2 for the mutant LMAN1 is weak or abolished by the binding of the anti-MCFD2 antibody. In addition, a Myc/63His-tagged recombinant form of wild-type LMAN1 could bind to D-mannose, but that of the mutant could not. The p.Trp67Ser mutation was located in the carbohydrate recognition domain (CRD), which is thought to participate in the selective binding of LMAN1 to the D-mannose of glycoproteins as well as the EF-motif of MCFD2. Taken together, it was suggested that the p.Trp67Ser mutation might affect the molecular chaperone function of LMAN1, impairing affinity for D-mannose as well as for MCFD2, which may be responsible for F5F8D in the patient. This is the first report of F5F8D caused by a qualitative defect of LMAN1 due to a missense mutation in LMAN1. Am. J. Hematol. 84:738-742, 2009. V V C 2009 WileyLiss, Inc. IntroductionCoagulation factor V (FV) and factor VIII (FVIII) are both essential in the blood coagulation cascade, as cofactors for the proteases factor X and factor IX, respectively. Combined deficiency of FV and FVIII (F5F8D) is an autosomal recessive bleeding disorder first described by Oeri et al. in 1954 [1], and a distinct clinical entity from chance co-inheritance of hemophilia A (FVIII deficiency) and parahemophilia (FV deficiency). F5F8D is extremely rare (1:2,000,000) in the general population [2], and characterized by a mild-to-moderate bleeding tendency manifested after surgical trauma, abortion, and delivery. Menorrhagia is also common, but hematuria and gastrointestinal bleeding are infrequent, and hemarthrosis is rare [3]. Generally, patients with F5F8D show plasma levels of FV and FVIII in the range of 5-30% of normal [4].Positional cloning has identified two genes, LMAN1 (lectin, mannose-binding, 1; also known as ERGIC-53) and MCFD2 (multiple coagulation factor deficiency gene 2), associated with F5F8D [4,5] LMAN1 is a Type-1 transmembrane protein that cycles between the endoplasmic reticulum (ER) and the ER-Golgi intermediate compartment (ERGIC) [6,7]. It contains a mannose-specific carbohydrate recognition domain (CRD) on the ER luminal side, and ER exit and retrieval motifs on the cytoplasmic side [8]. MCFD2 has an EF-hand domain that interacts with LMAN1 in a Ca 21 -dependent manner [5]. The LMAN1-MCFD2 protein complex functions as a cargo ...

show abstract

Transcription factor specificity protein 1 (Sp1) is the main regulator of nerve growth factor‐induced sphingosine kinase 1 gene expression of the rat pheochromocytoma cell line, PC12

Cited by 56 publications

References 45 publications

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

Molecular basis of antithrombin deficiency in four Japanese patients with antithrombin gene abnormalities including two novel mutations

Ewing's sarcoma fusion protein, EWS/Fli-1 and Fli-1 protein induce PLD2 but not PLD1 gene expression by binding to an ETS domain of 5′ promoter

A novel missense mutation causing abnormal LMAN1 in a Japanese patient with combined deficiency of factor V and factor VIII

Contact Info

Product

Resources

About