Identification and functional analysis of multiple murine myeloperoxidase (MPO) promoters and comparison with the human MPO promoter region

Zhao, Wei; Lu, J P; Regmi, Ajit; Austin, Garth E.

doi:10.1038/sj.leu.2400540

Cited by 23 publications

(16 citation statements)

References 3 publications

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“…Mouse-human species differences may be relevant in interpreting the role of phagocyte-inflicted injury using this model. MPO levels (per neutrophil) are fivefold to tenfold higher in the human than in the mouse (20,81), and MPO regulation, including active promoters (82) and inducers, may differ significantly. In contrast to human atheroma that readily stain for MPO and show clear evidence of chlorotyrosine (474 µmol/mol; ref.…”

mentioning

confidence: 99%

Increased atherosclerosis in myeloperoxidase-deficient mice

Brennan¹,

Anderson²,

Shih³

et al. 2001

J. Clin. Invest.

301

212

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mentioning

confidence: 99%

Increased atherosclerosis in myeloperoxidase-deficient mice

Brennan¹,

Anderson²,

Shih³

et al. 2001

J. Clin. Invest.

301

212

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“…This evidence implies the presence of an initiation site at bp −312, in agreement with the larger of the two bands seen with primer E. Together, these data support the existence of a second transcription start site for the MPO gene in HL-60 cells, approximately 312 bp upstream from the P1 site. While this novel initiation site appears to lie somewhat downstream from the approximate location we previously derived for our P2 promoter by a transient transfection approach, 9 that promoter was not accurately localized in our previous study. Furthermore, as shown below, no other initiation sites were observed between bp −312 and bp −900 by our primer extension experiments.…”

Section: Figurementioning

confidence: 74%

“…17,18 The location of this promoter corresponded closely with the MPO transcription initiation site described by Morishita et al 19 on the basis of S1 mapping experiments, and confirmed by Hashinaka et al 12 and Chang et al 14 However, more recently, we reported the existence of two additional human MPO promoters (termed 'P2' and 'P3'), whose activity could be demonstrated in transient transfection experiments. 9 Although we did not accurately localize the upstream promoters in these transfection studies, the P2 and P3 promoters appeared to be located approximately 500 bp and 1000 bp upstream from the P1 promoter, respectively. None of these three basal promoters, in themselves, showed tissue specificity or maturation stage specificity in transfection experiments, although addition of various enhancer elements conferred degrees of specificity upon the various promoters.…”

Section: Introductionmentioning

confidence: 74%

“…We also demonstrated the presence of three homologous promoters in the human MPO gene. 9 The importance of these diverse initiation sites in the murine MPO gene, and whether they are involved in differential regulation of MPO transcription in the mouse remain to be determined. However, the data which we provide in the present report suggest that the corresponding upstream initiation sites in the human MPO gene may play little role in physiologic regulation of MPO transcription.…”

Section: Discussionmentioning

confidence: 99%

“…Their findings suggested that at least three and possibly four initiation sites for MPO transcription are used in 32D Cl3 cells. To further evaluate the possible use of multiple MPO promoters in murine myeloid cells, we carried out transient transfection experiments designed to look for multiple promoters near the sites suggested by the data of Friedman et al Our results 9 demonstrated the existence of four distinct murine MPO promoters, which corresponded in location with the initiation sites previously demonstrated by sequence analysis of murine MPO cDNA clones.…”

Section: Introductionmentioning

confidence: 99%

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Functional activity of three distinct myeloperoxidase (MPO) promoters in human myeloid cells

Austin

2002

Leukemia

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The enzyme myeloperoxidase (MPO) is synthesized only in myeloid and monocytic cells, making it an important marker of the myeloid lineage. Transcription of the MPO gene is turned on early during the myeloblast stage of myeloid differentiation and is turned off when myeloid precursors are induced to differentiate along any one of a number of pathways. MPO transcripts show heterogeneity in size and sequence due, in part, to differential RNA splicing. We recently reported transfection studies which showed the presence of three distinct MPO promoters in the 5Ј-flanking region of the human MPO gene, suggesting that MPO transcription may also be regulated through the use of multiple promoters. We now report results of primer extension and RT-PCR experiments designed to determine if transcription of the human MPO gene is initiated at multiple promoter sites in vivo. MPO RNA obtained from myeloid cell lines was analyzed by primer extension using primers located at various sites between bp −1100 and bp +120 of the MPO gene. In addition, RT-PCR experiments were carried out using primer pairs located at intervals between bp −1000 and bp +2500 of the MPO gene. MPO transcripts were found to be initiated at three sites located about bp −920, bp −310, and bp +1 of the MPO gene, corresponding closely to our previously described P3, P2 and P1 promoters, respectively. Transcription initiated at the P1 site gave rise to large transcripts and showed the expected downregulation following induction of differentiation. On the other hand, transcripts initiated at the P3 and P2 sites did not show downregulation following induction of macrophage differentiation by TPA, and most did not appear to extend into the MPO coding region. Northern blot analysis of transcripts initiated at the P3 and P2 sites suggested that transcription at these sites was non-tissue-specific and indicated that many of these transcripts undergo premature termination. These results demonstrate that the MPO gene is transcribed in vivo primarily using the P1 promoter and that the low level of transcription occurring at the P2 and P3 sites is nonspecific and does not contribute significantly to physiologic regulation of MPO gene expression.

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Phagocytosis and peroxidase release by seabream (Sparus aurata L.) leucocytes in response to yeast cells

2003

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A flow cytometric method was adapted to evaluate phagocytosis by gilthead seabream leucocytes after incubation with yeast cells (Saccharomyces cerevisiae). Head-kidney leucocytes were incubated in vitro for different times in different proportions with heat-killed fluorescein isothiocyanate (FITC)-labeled yeast cells to study the kinetics of phagocytosis. Attached and internalized yeast cells were differentiated by quenching FITC-labeled S. cerevisiae with trypan blue dye. Only internalized cells kept their FITC fluorescence after quenching. Monocyte-macrophages and acidophilic granulocytes showed phagocytic activity, as demonstrated by transmission electron microscopy (TEM). From the ultrastructural features of the phagocytic process, it was observed that cytoplasmic granule membranes fused with the phagocyte membrane at the point where the yeast cell was attached to the phagocyte surface. This observation led us to adapt a colorimetric method to study peroxidase (myeloperoxidase and eosinophil peroxidase) release, since both are considered to be markers of the degranulation that occurs in seabream head-kidney leucocytes in response to yeast cells. Head-kidney leucocytes were incubated with calcium ionophore (CaI), phorbol myristate acetate (PMA), or yeast cells for different periods of time (0 -30 min) to study the kinetics of peroxidase release. The results obtained indicate that CaI and yeast cells, but not PMA, stimulate the degranulation (by about 44.51% and 21.04%, respectively, at 30 min) of seabream headkidney leucocytes. Anat Rec Part A 272A: 415-423, 2003.

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Identification and functional analysis of multiple murine myeloperoxidase (MPO) promoters and comparison with the human MPO promoter region

Abstract: and Friedman et al. 16 We also demonstrate that the human

Cited by 23 publications

References 3 publications

Increased atherosclerosis in myeloperoxidase-deficient mice

Increased atherosclerosis in myeloperoxidase-deficient mice

Functional activity of three distinct myeloperoxidase (MPO) promoters in human myeloid cells

Phagocytosis and peroxidase release by seabream (Sparus aurata L.) leucocytes in response to yeast cells

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