DOI: 10.1073/pnas.120571797. Down-regulation of iron regulatory protein 1 gene expression by nitric oxide

Down-regulation of iron regulatory protein 1 gene expression by nitric oxide

10.1073/pnas.120571797

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Iron regulatory protein 1 (IRP1) is an RNA binding protein that posttranscriptionally modulates the expression of mRNAs coding for proteins involved in iron metabolism. It has long been held that its RNA binding activity is regulated posttranslationally by the insertion/extrusion of a 4Fe-4S cluster, without changes in IRP1 levels. However, the question of a possible regulation of the expression of this protein has remained open. In the present study we analyzed the modulation of IRP1 expression in murine macrophages. We showed that activation by IFN-γ and/or lipopolysaccharide, which induces IRP1 RNA binding activity via nitric oxide (NO), results simultaneously in a reduction in IRP1 protein levels, as determined by Western blot analyses. IRP1 expression decreased time-dependently to about 40% of control levels after 16 h. Down-regulation of IRP1 protein levels was correlated with the amount of NO produced and was partially abolished by the NO synthase (NOS) inhibitorN-monomethyl-l-arginine. No changes in IRP1 levels could be detected in stimulated peritoneal macrophages from NOS2 knockout (NOS2−/−) mice, unlike wild-type mice. Converse modulation of IRP1 RNA binding activity and IRP1 levels could be reproduced by exogenous NO and also was observed in nonmacrophage cells cocultured with NO-producing macrophages. We also analyzed IRP1 mRNA levels by Northern blotting and found a decrease in IRP1 mRNA expression after stimulation with IFN-γ plus lipopolysaccharide, which was abrogated in the presence ofN-monomethyl-l-arginine. This is evidence that IRP1 is regulated by a physiological stimulus other than posttranslationally.

Bibliography

Oliveira, L., & Drapier, J.-C. (2000). Down-regulation of iron regulatory protein 1 gene expression by nitric oxide. Proceedings of the National Academy of Sciences, 97(12), 6550â6555.

Authors 2

Leonor Oliveira (first)
Jean-Claude Drapier (additional)

References 39 Referenced 42

10.1016/0092-8674(93)90046-S
10.1073/pnas.93.16.8175
10.1093/jn/128.12.2295
10.1074/jbc.270.37.21645
10.1002/j.1460-2075.1995.tb00219.x
10.1002/bies.950180909
10.1016/S0021-9258(18)35789-2
K Pantopoulos, N K Gray, M W Hentze RNA 1, 155–163 (1995). / RNA by Pantopoulos K (1995)
10.1073/pnas.89.24.11735
10.1096/fasebj.7.15.8262329
10.1073/pnas.89.24.11730
10.1002/j.1460-2075.1993.tb06038.x
10.1002/j.1460-2075.1993.tb06039.x
10.1074/jbc.274.1.516
10.1002/j.1460-2075.1995.tb07291.x
10.1074/jbc.271.12.7168
10.1074/jbc.273.13.7588
10.1074/jbc.273.16.9403
10.1126/science.2711187
10.1172/JCI112642
10.1016/S0076-6879(96)69006-5
10.1073/pnas.85.7.2171
10.1016/0092-8674(89)90851-9
10.1016/S0021-9258(19)74253-7
10.1007/s000180050355
10.1074/jbc.273.18.10857
10.1006/meth.1996.0426
10.1073/pnas.93.10.4925
10.1073/pnas.87.20.7958
10.1093/nar/20.1.33
10.1074/jbc.272.33.20340
10.1006/bbrc.1997.7930
10.1038/364450a0
10.1046/j.1471-4159.1994.62020496.x
A Habib, C Bernard, M Lebret, C Creminon, B Esposito, A Tedgui, J Maclouf J Immunol 158, 3845–3851 (1997). (10.4049/jimmunol.158.8.3845) / J Immunol by Habib A (1997)
10.1073/pnas.90.15.7240
10.1016/S0021-9258(18)41538-4
10.1074/jbc.272.40.25071
10.1016/S0378-1119(96)00329-0

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:35 a.m.)
Deposited	2 years, 4 months ago (April 22, 2023, 8:14 a.m.)
Indexed	4 months, 2 weeks ago (April 8, 2025, 5:38 p.m.)
Issued	25 years, 3 months ago (May 23, 2000)
Published	25 years, 3 months ago (May 23, 2000)
Published Online	25 years, 3 months ago (May 23, 2000)
Published Print	25 years, 2 months ago (June 6, 2000)

Funders 0

None

BibTeX

@article{Oliveira_2000, title={Down-regulation of iron regulatory protein 1 gene expression by nitric oxide}, volume={97}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.120571797}, DOI={10.1073/pnas.120571797}, number={12}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Oliveira, Leonor and Drapier, Jean-Claude}, year={2000}, month=may, pages={6550–6555} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        4,
        8
      ]
    ],
    "date-time": "2025-04-08T21:38:45Z",
    "timestamp": 1744148325940
  },
  "reference-count": 39,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "12",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2000,
        6,
        6
      ]
    ]
  },
  "abstract": "<jats:p>Iron regulatory protein 1 (IRP1) is an RNA binding protein that posttranscriptionally modulates the expression of mRNAs coding for proteins involved in iron metabolism. It has long been held that its RNA binding activity is regulated posttranslationally by the insertion/extrusion of a 4Fe-4S cluster, without changes in IRP1 levels. However, the question of a possible regulation of the expression of this protein has remained open. In the present study we analyzed the modulation of IRP1 expression in murine macrophages. We showed that activation by IFN-\u03b3 and/or lipopolysaccharide, which induces IRP1 RNA binding activity via nitric oxide (NO), results simultaneously in a reduction in IRP1 protein levels, as determined by Western blot analyses. IRP1 expression decreased time-dependently to about 40% of control levels after 16 h. Down-regulation of IRP1 protein levels was correlated with the amount of NO produced and was partially abolished by the NO synthase (NOS) inhibitor<jats:italic>N-</jats:italic>monomethyl-<jats:sc>l</jats:sc>-arginine. No changes in IRP1 levels could be detected in stimulated peritoneal macrophages from NOS2 knockout (NOS2<jats:sup>\u2212/\u2212</jats:sup>) mice, unlike wild-type mice. Converse modulation of IRP1 RNA binding activity and IRP1 levels could be reproduced by exogenous NO and also was observed in nonmacrophage cells cocultured with NO-producing macrophages. We also analyzed IRP1 mRNA levels by Northern blotting and found a decrease in IRP1 mRNA expression after stimulation with IFN-\u03b3 plus lipopolysaccharide, which was abrogated in the presence of<jats:italic>N-</jats:italic>monomethyl-<jats:sc>l</jats:sc>-arginine. This is evidence that IRP1 is regulated by a physiological stimulus other than posttranslationally.</jats:p>",
  "DOI": "10.1073/pnas.120571797",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:35:07Z",
    "timestamp": 1027694107000
  },
  "page": "6550-6555",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 42,
  "title": "Down-regulation of iron regulatory protein 1 gene expression by nitric oxide",
  "prefix": "10.1073",
  "volume": "97",
  "author": [
    {
      "given": "Leonor",
      "family": "Oliveira",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, Avenue de la Terrasse, 91190 Gif-sur-Yvette, France"
        }
      ]
    },
    {
      "given": "Jean-Claude",
      "family": "Drapier",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, Avenue de la Terrasse, 91190 Gif-sur-Yvette, France"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2000,
        5,
        23
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(93)90046-S"
    },
    {
      "key": "e_1_3_3_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.93.16.8175"
    },
    {
      "key": "e_1_3_3_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/jn/128.12.2295"
    },
    {
      "key": "e_1_3_3_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.37.21645"
    },
    {
      "key": "e_1_3_3_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1995.tb00219.x"
    },
    {
      "key": "e_1_3_3_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/bies.950180909"
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)35789-2"
    },
    {
      "key": "e_1_3_3_8_2",
      "first-page": "155",
      "volume": "1",
      "author": "Pantopoulos K",
      "year": "1995",
      "unstructured": "K Pantopoulos, N K Gray, M W Hentze RNA 1, 155\u2013163 (1995).",
      "journal-title": "RNA"
    },
    {
      "key": "e_1_3_3_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.89.24.11735"
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1096/fasebj.7.15.8262329"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.89.24.11730"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1993.tb06038.x"
    },
    {
      "key": "e_1_3_3_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1993.tb06039.x"
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.274.1.516"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1995.tb07291.x"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.12.7168"
    },
    {
      "key": "e_1_3_3_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.13.7588"
    },
    {
      "key": "e_1_3_3_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.16.9403"
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.2711187"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1172/JCI112642"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0076-6879(96)69006-5"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.85.7.2171"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(89)90851-9"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(19)74253-7"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s000180050355"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.18.10857"
    },
    {
      "key": "e_1_3_3_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/meth.1996.0426"
    },
    {
      "key": "e_1_3_3_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.93.10.4925"
    },
    {
      "key": "e_1_3_3_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.87.20.7958"
    },
    {
      "key": "e_1_3_3_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/20.1.33"
    },
    {
      "key": "e_1_3_3_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.33.20340"
    },
    {
      "key": "e_1_3_3_32_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/bbrc.1997.7930"
    },
    {
      "key": "e_1_3_3_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/364450a0"
    },
    {
      "key": "e_1_3_3_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1471-4159.1994.62020496.x"
    },
    {
      "key": "e_1_3_3_35_2",
      "doi-asserted-by": "crossref",
      "first-page": "3845",
      "DOI": "10.4049/jimmunol.158.8.3845",
      "volume": "158",
      "author": "Habib A",
      "year": "1997",
      "unstructured": "A Habib, C Bernard, M Lebret, C Creminon, B Esposito, A Tedgui, J Maclouf J Immunol 158, 3845\u20133851 (1997).",
      "journal-title": "J Immunol"
    },
    {
      "key": "e_1_3_3_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.90.15.7240"
    },
    {
      "key": "e_1_3_3_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)41538-4"
    },
    {
      "key": "e_1_3_3_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.40.25071"
    },
    {
      "key": "e_1_3_3_39_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0378-1119(96)00329-0"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.120571797",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        4,
        22
      ]
    ],
    "date-time": "2023-04-22T12:14:13Z",
    "timestamp": 1682165653000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.120571797"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2000,
        5,
        23
      ]
    ]
  },
  "references-count": 39,
  "journal-issue": {
    "issue": "12",
    "published-print": {
      "date-parts": [
        [
          2000,
          6,
          6
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.120571797"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.120571797",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2000,
        5,
        23
      ]
    ]
  },
  "assertion": [
    {
      "value": "1999-12-28",
      "order": 0,
      "name": "received",
      "label": "Received",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2000-05-23",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}