Deficient Transcription of Mouse Mast Cell Protease 4 Gene in Mutant Mice of mi/mi Genotype

The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of transcription factors (hereafter called MITF). We reported that expression of the mouse mast cell protease 5 (MMCP-5) and MMCP-6 genes were deficient in cultured mast cells (CMC) derived from mutant mice ofmi/mi genotype. Despite the reduced expression of both MMCP-5 and MMCP-6, their regulation mechanisms were different. Because MMCP-5 is a chymase and MMCP-6 a tryptase, there was a possibility that the difference in regulation mechanisms was associated with their different characteristics as proteases. We compared the regulation mechanisms of another chymase, MMCP-4, with those of MMCP-5 and MMCP-6. The expression of the MMCP-4 gene was also deficient in mi/mi CMC. The overexpression of the normal (+) MITF but not of mi-MITF normalized the poor expression of the MMCP-4 gene in mi/mi CMC, indicating the involvement of +-MITF in transactivation of the MMCP-4 gene. Although MMCP-4 is chymase as MMCP-5, the regulation of MMCP-4 expression was more similar to MMCP-6 than to MMCP-5. We also showed the deficient expression of granzyme B and cathepsin G genes inmi/mi CMC. Genes encoding granzyme B, cathepsin G, MMCP-4, and MMCP-5 are located on chromosome 14. Because all these genes showed deficient expression in mi/mi CMC, there is a possibility that MITF might regulate the expression of these genes through a locus control region.

Download Full-text

Abnormal Expression of Mouse Mast Cell Protease 5 Gene in Cultured Mast Cells Derived From Mutant mi/mi Mice

Blood ◽

10.1182/blood.v90.8.3057 ◽

1997 ◽

Vol 90 (8) ◽

pp. 3057-3066 ◽

Cited By ~ 73

Author(s):

Eiichi Morii ◽

Tomoko Jippo ◽

Tohru Tsujimura ◽

Koji Hashimoto ◽

Dae-Ki Kim ◽

...

Keyword(s):

Mast Cell ◽

Transcription Factors ◽

Mast Cells ◽

Promoter Region ◽

Leucine Zipper ◽

Consensus Sequence ◽

Mast Cell Protease ◽

Helix Loop Helix ◽

Regulation Mechanisms ◽

Mouse Mast Cell

Abstract Mast cells contain a lot of mast cell-specific proteases. We have reported that the expression of mouse mast cell protease 6 (MMCP-6) is remarkably reduced in both cultured mast cells (CMCs) and skin mast cells of mi/mi mutant mice. In the present study, we found that the expression of MMCP-5 was reduced in CMCs but not in skin mast cells of mi/mi mice, and we compared the regulation mechanisms of MMCP-5 with those of MMCP-6. The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of transcription factors (hereafter called MITF ). The consensus sequence recognized and bound by bHLH-Zip transcription factors is CANNTG. The overexpression of the normal (+) MITF but not of mi-MITF normalized the poor expression of the MMCP-5 gene in mi/mi CMCs, indicating the involvement of +-MITF in transactivation of the MMCP-5 gene. Although +-MITF directly bound CANNTG motifs in the promoter region of the MMCP-6 gene and transactivated it, the binding of +-MITF to the CAGTTG motif in the promoter region of the MMCP-5 gene was not detectable. The +-MITF appeared to regulate the transactivation of the MMCP-5 gene indirectly. Moreover, addition of stem cell factor to the medium normalized the expression of the MMCP-5 but not of the MMCP-6 gene in mi/mi CMCs. Despite the significant reduction of both MMCP-5 and MMCP-6 expressions in mi/mi CMCs, their regulation mechanisms appeared to be different.

Download Full-text

Regulation of mouse mast cell protease 6 gene expression by transcription factor encoded by the mi locus

Blood ◽

10.1182/blood.v88.7.2488.bloodjournal8872488 ◽

1996 ◽

Vol 88 (7) ◽

pp. 2488-2494 ◽

Cited By ~ 105

Author(s):

E Morii ◽

T Tsujimura ◽

T Jippo ◽

K Hashimoto ◽

K Takebayashi ◽

...

Keyword(s):

Mast Cell ◽

Transcription Factors ◽

Mast Cells ◽

Leucine Zipper ◽

Luciferase Activity ◽

Mast Cell Protease ◽

Helix Loop Helix ◽

3T3 Fibroblasts ◽

Mouse Mast Cell ◽

Nih 3T3

The mi locus of mice encodes a member of the basic-helix-loop-helix-leucine zipper (bHLH-Zip) protein family of transcription factors (hereafter called MITF). Because the expression of the mouse mast cell protease 6 (MMCP-6) gene is remarkably reduced in mast cells of mi/mi mutant mice, we investigated the effect of MITF on the transcription of the MMCP-6 gene. First, we introduced the normal (+) MITF cDNA into mi/mi cultured mast cells using the retroviral vector. Overexpression of +-MITF but not mi-MITF normalized the expression of the MMCP-6 gene, indicating the involvement of +-MITF in the MMCP-6 gene transactivation. Second, we analyzed the promoter of the MMCP-6 gene by the transient cotransfection assay. The luciferase construct under the control of the MMCP-6 promoter and the cDNA encoding +-MITF or mi-MITF were cotransfected into NIH/ 3T3 fibroblasts. The coexpression of +- MITF but not mi-MITF increased the luciferase activity 10-fold. We found a CACATG and a CATCTG motif in the MMCP-6 promoter, both of which are generally recognized by bHLH-Zip-type transcription factors. We also found a GACCTG motif that was strongly bound by +-MITF. These three motifs were necessary for the 10-fold transactivation ability of the MMCP-6 promoter by +-MITF. Mutations of each motif significantly reduced the transactivation, suggesting that +-MITF directly transactivated the MMCP-6 gene through these three motifs.

Download Full-text

Inhibitory effect of the transcription factor encoded by the mutant mi microphthalmia allele on transactivation of mouse mast cell protease 7 gene

Blood ◽

10.1182/blood.v97.3.645 ◽

2001 ◽

Vol 97 (3) ◽

pp. 645-651 ◽

Cited By ~ 34

Author(s):

Hideki Ogihara ◽

Eiichi Morii ◽

Dae-Ki Kim ◽

Keisuke Oboki ◽

Yukihiko Kitamura

Keyword(s):

Transcription Factor ◽

Mast Cell ◽

Inhibitory Effect ◽

Chromosome 17 ◽

Binding Motif ◽

Mast Cell Protease ◽

Coding Regions ◽

Helix Loop Helix ◽

Negative Effect ◽

Mouse Mast Cell

Abstract The transcription factor encoded by the mi locus (MITF) is a transcription factor of the basic-helix-loop-helix zipper protein family. Mice of mi/mi genotype express a normal amount of abnormal MITF, whereas mice oftg/tg genotype do not express any MITFs due to the transgene insertional mutation. The effect of normal (+) and mutant (mi) MITFs on the expression of mouse mast cell protease (MMCP) 6 and 7 was examined. Both MMCP-6 and MMCP-7 are tryptases, and their coding regions with high homology are closely located on chromosome 17. Both MMCP-6 and MMCP-7 genes are expressed in normal cultured mast cells (+/+ CMCs). Although the transcription of MMCP-6 gene was severely suppressed in bothmi/mi and tg/tg CMCs, that of MMCP-7 gene was severely suppressed only in mi/mi CMCs. The study identified the most significant segment for the transcription in the 5′ flanking region of MMCP-7 gene. Unexpectedly, no CANNTG motifs were found that are recognized and bound by +-MITF in this segment. Instead, there was an AP-1 binding motif, and binding of c-Jun to the AP-1 motif significantly enhanced the transcription of MMCP-7 gene. The complex formation of c-Jun with either +-MITF ormi-MITF was demonstrated. The binding of +-MITF to c-Jun enhanced the transactivation of MMCP-7 gene, and that ofmi-MITF suppressed the transactivation. Although the former complex was located only in the nucleus, the latter complex was predominantly found in the cytoplasm. The negative effect ofmi-MITF on the transcription of MMCP-7 gene appeared to be executed through the interaction with c-Jun.

Download Full-text

Faculty Opinions recommendation of Pivotal role of mouse mast cell protease 4 in the conversion and pressor properties of Big-endothelin-1.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718003609.793475629 ◽

2013 ◽

Author(s):

Richard L Stevens

Keyword(s):

Mast Cell ◽

Pivotal Role ◽

Endothelin 1 ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Mouse Mast Cell Protease-4-dependent production of ET-1 (1-31) and of plaque progression in an/INS; Apolipoprotein E knock-out mouse model of spontaneous atherosclerosis

Life Sciences ◽

10.1016/j.lfs.2013.12.192 ◽

2013 ◽

Vol 93 (25-26) ◽

pp. e57

Author(s):

Martin Houde ◽

Walid Semaan ◽

Louisane Desbiens ◽

Zhipeng You ◽

Adel G. Schwertani ◽

...

Keyword(s):

Mast Cell ◽

Mouse Model ◽

Apolipoprotein E ◽

Plaque Progression ◽

Knock Out ◽

Mast Cell Protease ◽

Mouse Mast Cell ◽

Knock Out Mouse

Download Full-text

Human mouse mast cell protease 7–like tryptase genes are pseudogenes

Journal of Allergy and Clinical Immunology ◽

10.1067/mai.2001.112130 ◽

2001 ◽

Vol 107 (2) ◽

pp. 315-321 ◽

Cited By ~ 14

Author(s):

Hae-Ki Min ◽

Naotomo Kambe ◽

Lawrence B. Schwartz

Keyword(s):

Mast Cell ◽

Mast Cell Protease ◽

Mouse Mast Cell

Download Full-text

Biochemical and immunological characterization of multiple glycoforms of mouse mast cell protease 1: comparison with an isolated murine serosal mast cell protease (MMCP-4)

Biochemical Journal ◽

10.1042/bj2940127 ◽

1993 ◽

Vol 294 (1) ◽

pp. 127-135 ◽

Cited By ~ 21

Author(s):

G F J Newlands ◽

D P Knox ◽

S R Pirie-Shepherd ◽

H R P Miller

Keyword(s):

Mast Cell ◽

Amino Acid ◽

Mast Cells ◽

Western Blotting ◽

Trichinella Spiralis ◽

Double Diffusion ◽

Terminal Amino Acid ◽

Mast Cell Protease ◽

Terminal Amino ◽

Mouse Mast Cell

Five highly soluble, chymotrypsin-like, neutral serine proteases, with molecular masses in the range 30-33 kDa, were isolated from Trichinella spiralis-infected mouse small intestine. These enzymes were closely related antigenically on Western blotting and by Ouchterlony double diffusion using a polyclonal, cross-absorbed, sheep antibody raised against mouse mast cell protease-1 (MMCP-1) and on the basis of N-terminal amino acid sequence analysis, were identified as variant forms of MMCP-1. Substrate and inhibitor analysis confirmed that the five variants (MMCP-1 A-E) had similar characteristics, although highly significant (P = 0.025 to P < 0.0001) variations in Km and kcat, were detected. Against human alpha 1-proteinase inhibitor the Ki for MMCP-1C (45 pM) was significantly (P < 0.0001) greater than those for the other proteases (0.76-2.2 pM). The differences in electrophoretic mobility are probably a result of variable glycosylation, since removal of N-linked carbohydrate produced a polypeptide of approx. 28 kDa in each case which was, like the native enzyme, immunoreactive on Western blotting. A much less soluble 28 kDa enzyme was isolated from serosal mast cells and identified as MMCP-4 by N-terminal amino acid sequencing. Like MMCP-1 it has chymotrypsin-like substrate specificities with activity at neutral pH. However, it was antigenically distinct from MMCP-1 and, using sheep anti-MMCP-1, was not detected on Western blotting or by Ouchterlony double diffusion, e.l.i.s.a. or immunohistochemistry. This last technique established that the MMCP-1 variants were uniquely present in enteric mast cells, thereby providing a highly selective means of distinguishing the mucosal and connective tissue mast cell subsets in the mouse.

Download Full-text

The Evolutionary History of the Chymase Locus -a Locus Encoding Several of the Major Hematopoietic Serine Proteases

International Journal of Molecular Sciences ◽

10.3390/ijms222010975 ◽

2021 ◽

Vol 22 (20) ◽

pp. 10975

Author(s):

Srinivas Akula ◽

Zhirong Fu ◽

Sara Wernersson ◽

Lars Hellman

Keyword(s):

Mast Cell ◽

T Cell ◽

Serine Proteases ◽

Phylogenetic Trees ◽

Granzyme B ◽

Large Family ◽

Cathepsin G ◽

Immune Functions ◽

New Class ◽

History Of

Several hematopoietic cells of the immune system store large amounts of proteases in cytoplasmic granules. The absolute majority of these proteases belong to the large family of chymotrypsin-related serine proteases. The chymase locus is one of four loci encoding these granule-associated serine proteases in mammals. The chymase locus encodes only four genes in primates, (1) the gene for a mast-cell-specific chymotryptic enzyme, the chymase; (2) a T-cell-expressed asp-ase, granzyme B; (3) a neutrophil-expressed chymotryptic enzyme, cathepsin G; and (4) a T-cell-expressed chymotryptic enzyme named granzyme H. Interestingly, this locus has experienced a number of quite dramatic expansions during mammalian evolution. This is illustrated by the very large number of functional protease genes found in the chymase locus of mice (15 genes) and rats (18 genes). A separate expansion has also occurred in ruminants, where we find a new class of protease genes, the duodenases, which are expressed in the intestinal region. In contrast, the opossum has only two functional genes in this locus, the mast cell (MC) chymase and granzyme B. This low number of genes may be the result of an inversion, which may have hindered unequal crossing over, a mechanism which may have been a major factor in the expansion within the rodent lineage. The chymase locus can be traced back to early tetrapods as genes that cluster with the mammalian genes in phylogenetic trees can be found in frogs, alligators and turtles, but appear to have been lost in birds. We here present the collected data concerning the evolution of this rapidly evolving locus, and how these changes in gene numbers and specificities may have affected the immune functions in the various tetrapod species.

Download Full-text