scholarly journals Systematic Method to Obtain Novel Genes That Are Regulated bymi Transcription Factor: Impaired Expression of Granzyme B and Tryptophan Hydroxylase in mi/mi Cultured Mast Cells

Blood ◽  
1998 ◽  
Vol 91 (9) ◽  
pp. 3210-3221 ◽  
Author(s):  
Akihiko Ito ◽  
Eiichi Morii ◽  
Kazutaka Maeyama ◽  
Tomoko Jippo ◽  
Dae-Ki Kim ◽  
...  
Keyword(s):  
Mast Cells ◽  
Leucine Zipper ◽  
Tryptophan Hydroxylase ◽  
Serotonin Synthesis ◽  
Systematic Method ◽  
Serotonin Content ◽  
Helix Loop Helix ◽  
New Genes ◽  
Rate Limiting ◽  
Leucine Zipper Protein

Abstract The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper protein family of transcription factors (hereafter called MITF). We have reported that the expression of several genes was impaired in cultured mast cells (CMCs) ofmi/mi genotype, and demonstrated the involvement of MITF in the transcription of these genes. To obtain new genes whose transcription may be regulated by MITF, we prepared a subtracted cDNA library using +/+ and mi/mi CMCs. We found two clones carrying the granzyme (Gr) B and tryptophan hydroxylase (TPH) cDNAs in the subtracted library. The expression of the Gr B and TPH genes decreased in mi/mi CMCs, and recovered to nearly normal level by the overexpression of normal (+) MITF but not of mutant (mi) MITF. The +-MITF bound three and one CANNTG motifs in the Gr B and TPH promoters, respectively, and transactivated these two genes, indicating the involvement of +-MITF in their expression. Because TPH is the rate-limiting enzyme for serotonin synthesis, we examined the serotonin content of +/+ and mi/mi CMCs. The serotonin content was significantly smaller in mi/mi CMCs than in +/+ CMCs. The introduction of +-MITF but not of mi-MITF normalized the serotonin content in mi/mi CMCs.

scholarly journals Systematic Method to Obtain Novel Genes That Are Regulated bymi Transcription Factor: Impaired Expression of Granzyme B and Tryptophan Hydroxylase in mi/mi Cultured Mast Cells

Blood ◽  
1998 ◽  
Vol 91 (9) ◽  
pp. 3210-3221 ◽  
Author(s):  
Akihiko Ito ◽  
Eiichi Morii ◽  
Kazutaka Maeyama ◽  
Tomoko Jippo ◽  
Dae-Ki Kim ◽  
...  
Keyword(s):  
Mast Cells ◽  
Leucine Zipper ◽  
Tryptophan Hydroxylase ◽  
Serotonin Synthesis ◽  
Systematic Method ◽  
Serotonin Content ◽  
Helix Loop Helix ◽  
New Genes ◽  
Rate Limiting ◽  
Leucine Zipper Protein

The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper protein family of transcription factors (hereafter called MITF). We have reported that the expression of several genes was impaired in cultured mast cells (CMCs) ofmi/mi genotype, and demonstrated the involvement of MITF in the transcription of these genes. To obtain new genes whose transcription may be regulated by MITF, we prepared a subtracted cDNA library using +/+ and mi/mi CMCs. We found two clones carrying the granzyme (Gr) B and tryptophan hydroxylase (TPH) cDNAs in the subtracted library. The expression of the Gr B and TPH genes decreased in mi/mi CMCs, and recovered to nearly normal level by the overexpression of normal (+) MITF but not of mutant (mi) MITF. The +-MITF bound three and one CANNTG motifs in the Gr B and TPH promoters, respectively, and transactivated these two genes, indicating the involvement of +-MITF in their expression. Because TPH is the rate-limiting enzyme for serotonin synthesis, we examined the serotonin content of +/+ and mi/mi CMCs. The serotonin content was significantly smaller in mi/mi CMCs than in +/+ CMCs. The introduction of +-MITF but not of mi-MITF normalized the serotonin content in mi/mi CMCs.

Importance of leucine zipper domain of mitranscription factor (MITF) for differentiation of mast cells demonstrated using mice/mice mutant mice of which MITF lacks the zipper domain

Blood ◽  
2001 ◽  
Vol 97 (7) ◽  
pp. 2038-2044 ◽  
Author(s):  
Eiichi Morii ◽  
Hideki Ogihara ◽  
Dae-Ki Kim ◽  
Akihiko Ito ◽  
Keisuke Oboki ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Mast Cells ◽  
Cytotoxic Activity ◽  
Leucine Zipper ◽  
Tryptophan Hydroxylase ◽  
Messenger Rnas ◽  
Serotonin Content ◽  
Helix Loop Helix ◽  
Inhibitory Function ◽  
Rate Limiting

Abstract The mi transcription factor (MITF) is a basic helix-loop-helix leucine zipper (bHLH-Zip) transcription factor that is important for the development of mast cells. Mast cells ofmi/mi genotype express normal amount of abnormal MITF (mi-MITF), whereas mast cells of tg/tg genotype do not express any MITFs. Mast cells of mi/mi mice show more severe abnormalities than those of tg/tg mice, indicating that the mi-MITF possesses the inhibitory function. The MITF encoded by the mice mutant allele (ce-MITF) lacks the Zip domain. We examined the importance of the Zip domain usingmice/micemice. The amounts of c-kit, granzyme B (Gr B), and tryptophan hydroxylase (TPH) messenger RNAs decreased in mast cells ofmice/mice mice to levels comparable to those of tg/tg mice, and the amounts were intermediate between those of +/+ mice and those of mi/mi mice. Gr B mediates the cytotoxic activity of mast cells, and TPH is a rate-limiting enzyme for the synthesis of serotonin. The cytotoxic activity and serotonin content ofmice/mice mast cells were comparable to those of tg/tg mast cells and were significantly higher than those of mi/mi mast cells. The phenotype of mice/mice mast cells was similar to that of tg/tg mast cells rather than to that of mi/mi mast cells, suggesting that thece-MITF had no functions. The Zip domain of MITF appeared to be important for the development of mast cells.

Inhibitory Effect of the Transcription Factor Encoded by themi Mutant Allele in Cultured Mast Cells of Mice

Blood ◽  
1999 ◽  
Vol 93 (4) ◽  
pp. 1189-1196 ◽  
Author(s):  
Akihiko Ito ◽  
Eiichi Morii ◽  
Dae-Ki Kim ◽  
Tatsuki R. Kataoka ◽  
Tomoko Jippo ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Mast Cells ◽  
Leucine Zipper ◽  
Tryptophan Hydroxylase ◽  
Cdna Probe ◽  
Inhibitory Effect ◽  
Northern Analysis ◽  
Helix Loop Helix ◽  
Genes Encoding ◽  
Mouse Mast Cell

The mi locus of mice encodes a transcription factor of the basic-helix-loop-helix-leucine zipper protein family (MITF). The MITF encoded by the mutant mi allele (mi-MITF) deletes 1 of 4 consecutive arginines in the basic domain. The mice of mi/migenotype express mi-MITF, whereas the mice of tg/tggenotype have a transgene at the 5′ flanking region of themi gene and do not express any MITF. To investigate the function of mi-MITF in cultured mast cells (CMCs), we took two approaches. First, mRNA obtained from mi/mi CMCs ortg/tg CMCs was subtracted from complementary (c) DNA library of normal (+/+) CMCs, and the (+/+-mi/mi) and (+/+-tg/tg) subtraction libraries were obtained. When the number of clones that hybridized more efficiently with +/+ CMC cDNA probe than with mi/mi or tg/tg CMC cDNA probe was compared using Southern analysis, the number was larger in the (+/+-mi/mi) library than in the (+/+-tg/tg) library. Second, we compared mRNA expression of six genes betweenmi/mi and tg/tg CMCs by Northern analysis. The transcription of three genes encoding mouse mast cell proteases was impaired in both mi/mi and tg/tg CMCs. On the other hand, the transcription of three genes encoding c-kit receptor, tryptophan hydroxylase, and granzyme B was markedly reduced inmi/mi CMCs, but the reduction was significantly smaller intg/tg CMCs. These results indicated the inhibitory effect ofmi-MITF on the transactivation of particular genes in CMCs.

Impaired Expression of Integrin -4 Subunit in Cultured Mast Cells Derived From Mutant Mice of mi/mi Genotype

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 1973-1980 ◽  
Author(s):  
Dae-Ki Kim ◽  
Eiichi Morii ◽  
Hideki Ogihara ◽  
Koji Hashimoto ◽  
Kenji Oritani ◽  
...  
Keyword(s):  
Mast Cells ◽  
Cell Adhesion Molecule ◽  
Leucine Zipper ◽  
Subunit Gene ◽  
Vascular Cell Adhesion ◽  
Gene Encoding ◽  
Helix Loop Helix ◽  
American Society ◽  
Cell Adhesion Molecule 1 ◽  
Leucine Zipper Protein

Abstract The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper protein family of transcription factors (hereafter called MITF). We have reported that expression of several genes was impaired in cultured mast cells (CMCs) ofmi/mi mice due to a defective transactivation ability of mutant MITF (mi-MITF). Because attachment of mi/mi CMCs to fibroblasts is impaired, we examined the expression of integrin genes in mi/mi CMCs in the present study. Among the integrin genes examined, the expression of integrin 4 subunit was barely detectable in mi/mi CMCs, and the 4 protein was not detected by flow cytometry either. The specific adhesion to vascular cell adhesion molecule-1 (VCAM-1), the ligand for 4 subunit, was observed in +/+ CMCs but not in mi/mi CMCs, indicating that the expression of integrin 4 subunit at a functional level did not occur in mi/mi CMCs. In the promoter region of the 4 subunit gene, there was a CACTTG motif to which normal MITF (+- MITF) bound. The coexpression of +-MITF but not of mi-MITF transactivated the promoter of the 4 subunit gene. The deletion or mutation of the CACTTG motif abolished the transactivation by +-MITF, suggesting that +-MITF directly transactivated the gene encoding 4 subunit of integrin. © 1998 by The American Society of Hematology.

Inhibitory Effect of the Transcription Factor Encoded by themi Mutant Allele in Cultured Mast Cells of Mice

Blood ◽  
1999 ◽  
Vol 93 (4) ◽  
pp. 1189-1196 ◽  
Author(s):  
Akihiko Ito ◽  
Eiichi Morii ◽  
Dae-Ki Kim ◽  
Tatsuki R. Kataoka ◽  
Tomoko Jippo ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Mast Cells ◽  
Leucine Zipper ◽  
Tryptophan Hydroxylase ◽  
Cdna Probe ◽  
Inhibitory Effect ◽  
Northern Analysis ◽  
Helix Loop Helix ◽  
Genes Encoding ◽  
Mouse Mast Cell

Abstract The mi locus of mice encodes a transcription factor of the basic-helix-loop-helix-leucine zipper protein family (MITF). The MITF encoded by the mutant mi allele (mi-MITF) deletes 1 of 4 consecutive arginines in the basic domain. The mice of mi/migenotype express mi-MITF, whereas the mice of tg/tggenotype have a transgene at the 5′ flanking region of themi gene and do not express any MITF. To investigate the function of mi-MITF in cultured mast cells (CMCs), we took two approaches. First, mRNA obtained from mi/mi CMCs ortg/tg CMCs was subtracted from complementary (c) DNA library of normal (+/+) CMCs, and the (+/+-mi/mi) and (+/+-tg/tg) subtraction libraries were obtained. When the number of clones that hybridized more efficiently with +/+ CMC cDNA probe than with mi/mi or tg/tg CMC cDNA probe was compared using Southern analysis, the number was larger in the (+/+-mi/mi) library than in the (+/+-tg/tg) library. Second, we compared mRNA expression of six genes betweenmi/mi and tg/tg CMCs by Northern analysis. The transcription of three genes encoding mouse mast cell proteases was impaired in both mi/mi and tg/tg CMCs. On the other hand, the transcription of three genes encoding c-kit receptor, tryptophan hydroxylase, and granzyme B was markedly reduced inmi/mi CMCs, but the reduction was significantly smaller intg/tg CMCs. These results indicated the inhibitory effect ofmi-MITF on the transactivation of particular genes in CMCs.

Impaired Expression of Integrin -4 Subunit in Cultured Mast Cells Derived From Mutant Mice of mi/mi Genotype

Blood ◽  
1998 ◽  
Vol 92 (6) ◽  
pp. 1973-1980 ◽  
Author(s):  
Dae-Ki Kim ◽  
Eiichi Morii ◽  
Hideki Ogihara ◽  
Koji Hashimoto ◽  
Kenji Oritani ◽  
...  
Keyword(s):  
Mast Cells ◽  
Cell Adhesion Molecule ◽  
Leucine Zipper ◽  
Subunit Gene ◽  
Vascular Cell Adhesion ◽  
Gene Encoding ◽  
Helix Loop Helix ◽  
American Society ◽  
Cell Adhesion Molecule 1 ◽  
Leucine Zipper Protein

The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper protein family of transcription factors (hereafter called MITF). We have reported that expression of several genes was impaired in cultured mast cells (CMCs) ofmi/mi mice due to a defective transactivation ability of mutant MITF (mi-MITF). Because attachment of mi/mi CMCs to fibroblasts is impaired, we examined the expression of integrin genes in mi/mi CMCs in the present study. Among the integrin genes examined, the expression of integrin 4 subunit was barely detectable in mi/mi CMCs, and the 4 protein was not detected by flow cytometry either. The specific adhesion to vascular cell adhesion molecule-1 (VCAM-1), the ligand for 4 subunit, was observed in +/+ CMCs but not in mi/mi CMCs, indicating that the expression of integrin 4 subunit at a functional level did not occur in mi/mi CMCs. In the promoter region of the 4 subunit gene, there was a CACTTG motif to which normal MITF (+- MITF) bound. The coexpression of +-MITF but not of mi-MITF transactivated the promoter of the 4 subunit gene. The deletion or mutation of the CACTTG motif abolished the transactivation by +-MITF, suggesting that +-MITF directly transactivated the gene encoding 4 subunit of integrin. © 1998 by The American Society of Hematology.

Additive effect of mouse genetic background and mutation ofMITF gene on decrease of skin mast cells

Blood ◽  
2003 ◽  
Vol 101 (4) ◽  
pp. 1344-1350 ◽  
Author(s):  
Eiichi Morii ◽  
Keisuke Oboki ◽  
Tomoko Jippo ◽  
Yukihiko Kitamura
Keyword(s):  
Transcription Factor ◽  
Mast Cells ◽  
Receptor Tyrosine Kinase ◽  
Genetic Background ◽  
Leucine Zipper ◽  
Basic Helix Loop Helix ◽  
Kit Ligand ◽  
Helix Loop Helix ◽  
Kit Receptor ◽  
Mouse Genetic

The mi transcription factor (MITF) is a basic-helix-loop-helix leucine zipper transcription factor and is encoded by mi locus. The mi/mi mutant mice showed a significant decrease of skin mast cells in C57BL/6 (B6) genetic background but not in WB genetic background. Kit ligand (KitL) is the most important growth factor for development of mast cells, and the decrease of skin mast cells in B6-mi/mi mice was attributable to the reduced expression of c-kit receptor tyrosine kinase (KIT) that is a receptor for KitL. However, the expression level of KIT in WB-mi/mi mast cells was comparable with that of B6-mi/mi mast cells, suggesting that a factor compensating the reduced expression of KIT was present in WB-mi/mi mice. By linkage analysis, such a factor was mapped on chromosome 10. The mapped position was closely located to the KitL locus. Two alternative spliced forms are known in KitL mRNA: KL-1 and KL-2. Soluble KitL, which is important for development of skin mast cells, is produced more efficiently from KL-1 mRNA than from KL-2 mRNA. The KL-1/KL-2 ratio was higher in WB-mi/mi than in B6-mi/mi mice, suggesting that the larger amount of soluble KitL may compensate for the reduced expression of KIT in WB-mi/mi mice.

scholarly journals Poor response of cultured mast cells derived from mi/mi mutant mice to nerve growth factor

Blood ◽  
1994 ◽  
Vol 84 (9) ◽  
pp. 2977-2983 ◽  
Author(s):  
T Jippo ◽  
H Ushio ◽  
S Hirota ◽  
H Mizuno ◽  
A Yamatodani ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Mast Cells ◽  
Leucine Zipper ◽  
Poor Response ◽  
Mutant Mice ◽  
Nerve Growth ◽  
Helix Loop Helix ◽  
Suboptimal Dose ◽  
And Control

Decreased numbers of mast cells and abnormalities in the phenotype of mast cells are observed in the skin of mi/mi mutant mice. Recently, the mi locus was identified to encode a novel member of the basic-helix- loop-helix-leucine zipper protein family of transcription factors. Since nerve growth factor (NGF) has been reported to influence the proliferation and the phenotype of cultured mast cells (CMCs), we compared the effect of NGF between mi/mi and control normal (+/+) CMCs. Addition of NGF to the suboptimal dose of recombinant murine interleukin-3 (rmIL-3) increased the plating efficiency of +/+ CMCs, but not of mi/mi CMCs. Although +/+ CMCs were berberine sulfate- negative when cultured with rmIL-3 alone, +/+ CMCs became berberine sulfate-positive when cultured in the presence of both rmIL-3 and NGF, which suggests increased heparin content. In contrast, NGF did not influence the phenotype of mi/mi CMCs. +/+ CMCs significantly bound 125I-NGF, but mi/mi CMCs did not, which suggests a defect of NGF receptors in mi/mi CMCs. Both p75 and p140 molecules are known to be involved in the formation of NGF receptors. Although the expression of p140 messenger (m)RNA was comparable between +/+ and mi/mi CMCs, the expression of p75 mRNA was significantly lower in mi/mi CMCs than in +/+ CMCs. Taken together, the poor response of mi/mi CMCs to NGF appeared to be attributable to the impaired transcription of the p75 gene.

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