Structure and Localization of the Mouse Prolyl Oligopeptidase Gene

Kimura, Atsushi; Yoshida, Itsuro; Takagi, Nobuo; Takahashi, Takayuki

doi:10.1074/jbc.274.34.24047

Cited by 20 publications

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References 60 publications

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“…After adding 46 l of 65 mM Tris-HCl (pH 8.3), 4.3 mM MgCl 2 , 15 mM dithiothreitol, and 0.72 mM dNTP mixture, 200 units of Superscript II reverse transcriptase (Life Technologies, Inc.) was introduced into the reaction mixture, which was incubated at 42°C for 60 min. The reactions were phenol-extracted, ethanol-precipitated, and electrophoresed in a denaturing 6% polyacrylamide sequencing gel along with a sequence ladder generated with the unlabeled primers using [␣- Genomic Southern Blotting Analysis-Mouse genomic DNA was extracted as described previously (10). 5 g of the genomic DNA was completely digested with EcoRI, HindIII, BamHI, and XhoI.…”

Section: The Nucleotide Sequence(s) Reported In This Paper Has Been Smentioning

confidence: 99%

Molecular and Biochemical Characterization of a Serine Proteinase Predominantly Expressed in the Medulla Oblongata and Cerebellar White Matter of Mouse Brain

Matsui¹,

Kimura²,

Yamashiki³

et al. 2000

Journal of Biological Chemistry

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A full-length cDNA clone of a serine proteinase, mouse brain serine proteinase (mBSP), was isolated from a mouse brain cDNA library. mBSP, which has been recently reported to be expressed in the hair follicles of nude mice, is most similar (88% identical) in sequence to rat myelencephalon-specific protease. The mBSP mRNA was steadily expressed in the brain of adult mice with a transient expression in the early fetal stage during development. The genomic structure of the mouse gene for mBSP was determined. The gene, which is mapped to chromosome 7B4-B5, is about 7.4 kilobases in size and contains 7 exons. Interestingly, the 5-untranslated region of the mBSP gene was interrupted by two introns. In situ hybridization analyses revealed that mBSP is expressed in the white matter of the cerebellum, medulla oblongata, and capsula interna and capsula interna pars retrolenticularis of mouse brain. Further, mBSP was immunolocalized to the neuroglial cells in the white matter of the cerebellum. Recombinant mBSP was produced in the bacterial expression system and activated by lysyl endopeptidase digestion, and the activated enzyme was purified for characterization. The enzyme showed amidolytic activities preferentially cleaving Arg-X bonds when 4-methylcoumaryl-7-amidecontaining peptide substrates were used. Typical serine proteinase inhibitors, such as diisopropyl fluorophosphates, phenylmethanesulfonyl fluoride, soybean trypsin inhibitor, aprotinin, leupeptin, antipain, and benzamidine, strongly inhibited the enzyme activity. The recombinant mBSP effectively hydrolyzed fibronectin and gelatin, but not laminin, collagens I and IV, or elastin. These results suggest that mBSP plays an important role in association with the function of the adult mouse brain.

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Section: The Nucleotide Sequence(s) Reported In This Paper Has Been Smentioning

confidence: 99%

Molecular and Biochemical Characterization of a Serine Proteinase Predominantly Expressed in the Medulla Oblongata and Cerebellar White Matter of Mouse Brain

Matsui¹,

Kimura²,

Yamashiki³

et al. 2000

Journal of Biological Chemistry

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“…A 914-bp mouse POP promoter (an EcoRI-BamHI fragment) was already subcloned into the upstream of the EGFP gene of a pEGFP-1 vector (Clontech Laboratories Inc, Palo Alto, CA, USA) (Kimura et al, 1999). By digesting it with XhoI and AflII restriction enzymes, the transgene composed of the POP promoter, the EGFP gene, and the SV40 poly A signal was isolated from the vector sequence.…”

Section: Generation Maintenance and Genotyping Of Transgenic Micementioning

confidence: 99%

“…This promoter has been already proved to have sufficient activity to drive the transcription of reporter genes in vitro (Kimura et al, 1999;Matsubara et al, 2010). By injecting the transgene into fertilized eggs, we obtained nine transgenic founders.…”

Section: Establishment Of Transgenic Lines and Determination Of Theirmentioning

confidence: 99%

“…A placenta-specific transcription factor, AP-2, could be a candidate because it was expressed in the same manner as POP (Sapin et al, 2000;Shi and Kellems, 1998) and the POP promoter had several AP-2 binding sites (Kimura et al, 1999). This time, we searched for the candidate AP-2 binding sites again by using the TRANSFAC software (http://www.gene-regulation.com/pub/databases.html) and found nine sites in the 914-bp POP promoter, one of which was the sequence for AP-2 (Fig.…”

Section: The Binding Of Ap-2 To the Pop Promotermentioning

confidence: 99%

“…The enhancer is the sequence to which activator proteins bind to increase the rate of transcription (Bulger and Groudine, 2011), and the insulator is a sequence to protect a transgene from position effect (Gaszner and Felsenfeld, 2006;Yang and Corces, 2011). In case of the POP gene, we determined a minimum promoter to drive its transcription in vitro (Kimura et al, 1999) and identified a CpG island in the genebody to be an enhancer candidate in the ovarian granulosa cell (Matsubara et al, 2010), but mechanisms for its transcriptional regulation are largely unknown.…”

Section: Introductionmentioning

confidence: 99%

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A 914-bp promoter is sufficient to reproduce the endogenous prolyl oligopeptidase gene localization in the mouse placenta if not subject to position effect

Matsubara

Maruyama

Kimura

2013

Gene

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Prolyl oligopeptidase (POP) is a widely distributed multifunctional protein which has an endopeptidase activity to cleave a -Pro-X- peptide bond. In spite of numerous studies about POP, the mechanism by which its transcription is controlled has not been well investigated. Here we generated transgenic mice bearing a transgene which contained a 914-bp POP gene promoter linked to the enhanced green fluorescent protein (EGFP) gene to assess the in vivo promoter activity. We established six transgenic lines with different copy numbers, but no EGFP signal was observed in four lines due to a high level of DNA methylation, which suggested that the transgene was subject to position effect. However, in the other two lines, we detected the EGFP expression in many tissues, and its placental localization showed a similar change to POP. A strong EGFP signal was observed in the junctional and labyrinthine zones of E10.5-E12.5 placentas and in the junctional zone and the maternal decidua after that. This placental gene activation might be attributed to AP-2 gamma because we detected its binding to the POP promoter. In contrast, we did not obtain any evidence that EGFP was expressed in a similar pattern compared with POP in the ovary. The current data demonstrated that the 914-bp promoter had sufficient activity to reproduce the POP localization in the placenta if it was not subject to position effect and suggest that the regulatory mechanism of the POP gene expression differs between tissues.(C) 2013 Elsevier B.V. All rights reserved

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cDNA cloning of rat prolyl oligopeptidase and its expression in the ovary during the estrous cycle

Kimura

Takahashi

2000

J. Exp. Zool.

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A cDNA for rat prolyl oligopeptidase was cloned which contained an open reading frame of 2,130 nucleotides encoding a protein of 710 amino acids. The deduced amino acid sequence is around 95% homologous to other mammalian prolyl oligopeptidases and about 40% to bacterial prolyl oligopeptidases. The recombinant prolyl oligopeptidase generated in E. coli was purified and its properties were examined. The substrate specificity and the susceptibility to proteinase inhibitors were similar to those of the native enzyme. Northern blot analysis showed wide expression of the prolyl oligopeptidase gene. Using ovaries from hormone‐treated rats, it was found that both the mRNA expression and enzyme activity increased in the luteal phase. These findings suggest the involvement of prolyl oligopeptidase in events associated with corpus luteum formation and/or luteal regression. J. Exp. Zool. 286:656–665, 2000. © 2000 Wiley‐Liss, Inc.

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Structure and Localization of the Mouse Prolyl Oligopeptidase Gene

Cited by 20 publications

References 60 publications

Molecular and Biochemical Characterization of a Serine Proteinase Predominantly Expressed in the Medulla Oblongata and Cerebellar White Matter of Mouse Brain

Molecular and Biochemical Characterization of a Serine Proteinase Predominantly Expressed in the Medulla Oblongata and Cerebellar White Matter of Mouse Brain

A 914-bp promoter is sufficient to reproduce the endogenous prolyl oligopeptidase gene localization in the mouse placenta if not subject to position effect

cDNA cloning of rat prolyl oligopeptidase and its expression in the ovary during the estrous cycle

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