CoChat
Sign Up
Cited by 194Metalloenzymes and iron-sulfur proteinsEnzyme Structure and FunctionInorganic Chemistry and Materials

Subcellular compartmentalization of human Nfu, an iron-sulfur cluster scaffold protein, and its ability to assemble a [4Fe-4S] cluster

Open access
Tong, Wing-Hang, Jameson, Guy N. L., Huynh, Boi Hanh, Rouault, Tracey A.·2003·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.1732541100PMC PMC187839
Open this paper in CoChat — free account

Abstract

Iron-sulfur (Fe-S) clusters serve as cofactors in many proteins that have important redox, catalytic, and regulatory functions. In bacteria, biogenesis of Fe-S clusters is mediated by multiple gene products encoded by the isc and nif operons. In particular, genetic and biochemical studies suggest that IscU, Nfu, and IscA function as scaffold proteins for assembly and delivery of rudimentary Fe-S clusters to target proteins. Here we report the characterization of human Nfu. A combination of biochemical and spectroscopic techniques, including UV-visible absorption and 57Fe Mössbauer spectroscopies, have been used to investigate the ability of purified human Nfu to assemble Fe-S clusters. The results suggest that Nfu can assemble approximately one labile [4Fe-4S] cluster per two Nfu monomers, and support the proposal that Nfu is an alternative scaffold protein for assembly of clusters that are subsequently used for maturation of targeted Fe-S proteins. Analyses of genomic DNA, transcripts, and translation products indicate that alternative splicing of a common pre-mRNA results in synthesis of two Nfu isoforms with distinct subcellular localizations. Isoform I is localized in the mitochondria, whereas isoform II is present in the cytosol and the nucleus. These results, together with previous reports of subcellular distributions of isoforms of human IscS and IscU in mitochondria, cytosol, and nucleus suggest that the Fe-S cluster assembly machineries are compartmentalized in higher eukaryotes.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1073/pnas.1732541100

Cites

54 of 54 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Mitochondrial iron is required for the synthesis of Fe-S clusters, and their biogenesis is thought to be one of the most important functions of mitochondria.

    “Mitochondrial iron is also required for the synthesis of Fe-S clusters and indeed their biogenesis is thought to be one of the most important functions of the mitochondria—even though some Fe-S assembly can occur in the cytosol and nucleus (Tong, 2000; Tong et al., 2003; Tong and Rouault, 2006).”

    From The Role of Iron Regulation in Immunometabolism and Immune-Related Disease (2019) · cites this paper as Tong et al., 2003

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 54

  1. [1]Beinert, H., Holm, R. H. & Münck, E. (1997) Science 277, 653–659. [DOI] [PubMed] [Google Scholar]
  2. [2]Jacobson, M. R., Brigle, K. E., Bennett, L. T., Setterquist, R. A., Wilson, M. S., Cash, V. L., Beynon, J., Newton, W. E. & Dean, D. R. (1989) J. Bacteriol. 171, 1017–1027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. [3]Jacobson, M. R., Cash, V. L., Weiss, M. C., Laird, N. F., Newton, W. E. & Dean, D. R. (1989) Mol. Gen. Genet. 219, 49–57. [DOI] [PubMed] [Google Scholar]
  4. [4]Zheng, L., White, R. H., Cash, V. L., Jack, R. F. & Dean, D. R. (1993) Proc. Natl. Acad. Sci. USA 90, 2754–2758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. [5]Fu, W., Jack, R. F., Morgan, T. V., Dean, D. R. & Johnson, M. K. (1994) Biochemistry 33, 13455–13463. [DOI] [PubMed] [Google Scholar]
  6. [6]Yuvaniyama, P., Agar, J. N., Cash, V. L., Johnson, M. K. & Dean, D. R. (2000) Proc. Natl. Acad. Sci. USA 97, 599–604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. [7]Zheng, L., Cash, V. L., Flint, D. H. & Dean, D. R. (1998) J. Biol. Chem. 273, 13264–13272. [DOI] [PubMed] [Google Scholar]
  8. [8]Tokumoto, U. & Takahashi, Y. (2001) J. Biochem. (Tokyo) 130, 63–71. [DOI] [PubMed] [Google Scholar]
  9. [9]Schwartz, C. J., Giel, J. L., Patschkowski, T., Luther, C., Ruzicka, F. J., Beinert, H. & Kiley, P. J. (2001) Proc. Natl. Acad. Sci. USA 98, 14895–14900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. [10]Agar, J. N., Zheng, L., Cash, V. L., Dean, D. R. & Johnson, M. K. (2000) J. Am. Chem. Soc. 122, 2136–2137. [Google Scholar]
  11. [11]Ollagnier-de-Choudens, S., Mattioli, T., Takahashi, Y. & Fontecave, M. (2001) J. Biol. Chem. 276, 22604–22607. [DOI] [PubMed] [Google Scholar]
  12. [12]Krebs, C., Agar, J. N., Smith, A. D., Frazzon, J., Dean, D. R., Huynh, B. H. & Johnson, M. K. (2001) Biochemistry 40, 14069–14080. [DOI] [PubMed] [Google Scholar]
  13. [13]Hoff, K. G., Silberg, J. J. & Vickery, L. E. (2000) Proc. Natl. Acad. Sci. USA 97, 7790–7795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. [14]Zheng, L., White, R. H., Cash, V. L. & Dean, D. R. (1994) Biochemistry 33, 4714–4720. [DOI] [PubMed] [Google Scholar]
  15. [15]Urbina, H. D., Silberg, J. J., Hoff, K. G. & Vickery, L. E. (2001) J. Biol. Chem. 276, 44521–44526. [DOI] [PubMed] [Google Scholar]
  16. [16]Smith, A. D., Agar, J. N., Johnson, K. A., Frazzon, J., Amster, I. J., Dean, D. R. & Johnson, M. K. (2001) J. Am. Chem. Soc. 123, 11103–11104. [DOI] [PubMed] [Google Scholar]
  17. [17]Agar, J. N., Krebs, C., Frazzon, J., Huynh, B. H., Dean, D. R. & Johnson, M. K. (2000) Biochemistry 39, 7856–7862. [DOI] [PubMed] [Google Scholar]
  18. [18]Nakai, M., Nishio, K., Morimoto, K., Yabe, T. & Kikuchi, S. (2002) Recent Res. Dev. Proteins 1, 1–11. [Google Scholar]
  19. [19]Ouzounis, C., Bork, P. & Sander, C. (1994) Trends Biochem. Sci. 19, 199–200. [DOI] [PubMed] [Google Scholar]
  20. [20]Agar, J. N., Yuvaniyama, P., Jack, R. F., Cash, V. L., Smith, A. D., Dean, D. R. & Johnson, M. K. (2000) J. Biol. Inorg. Chem. 5, 167–177. [DOI] [PubMed] [Google Scholar]
  21. [21]Garland, S. A., Hoff, K., Vickery, L. E. & Culotta, V. C. (1999) J. Mol. Biol. 294, 897–907. [DOI] [PubMed] [Google Scholar]
  22. [22]Schilke, B., Voisine, C., Beinert, H. & Craig, E. (1999) Proc. Natl. Acad. Sci. USA 96, 10206–10211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. [23]Nakamura, Y., Kaneko, T., Hirosawa, M., Miyajima, N. & Taba, S. (1998) Nucleic Acids Res. 26, 63–67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. [24]Nishio, K. & Nakai, M. (2000) J. Biol. Chem. 275, 22615–22618. [DOI] [PubMed] [Google Scholar]
  25. [25]Lorain, S., Lecluse, Y., Scamps, C., Mattei, M.-G. & Lipinsku, M. (2001) Biochim. Biophys. Acta 1517, 376–383. [DOI] [PubMed] [Google Scholar]
  26. [26]Land, T. & Rouault, T. A. (1998) Mol. Cell 2, 807–815. [DOI] [PubMed] [Google Scholar]
  27. [27]Tong, W.-H. & Rouault, T. (2000) EMBO J. 19, 5692–5700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. [28]Stookey, L. L. (1970) Anal. Chem. 42, 779–781. [Google Scholar]
  29. [29]Nakai, K. & Horton, P. (1999) Trends Biochem. Sci. 24, 34–35. [DOI] [PubMed] [Google Scholar]
  30. [30]Gavel, Y. & von Heijne, G. (1990) Protein Eng. 4, 33–37. [DOI] [PubMed] [Google Scholar]
  31. [31]Pfanner, N., Craig, E. A. & Honlinger, A. (1997) Annu. Rev. Cell Dev. Biol. 13, 25–51. [DOI] [PubMed] [Google Scholar]
  32. [32]Orme-Johnson, W. H. & Orme-Johnson, N. R. (1982) in Iron-Sulfur Proteins, ed. Spiro, T. G. (Wiley, New York), Vol. 4, pp. 67–146. [Google Scholar]
  33. [33]Middleton, P., Dickson, D. P. E., Johnson, C. E. & Rush, J. D. (1978) Eur. J. Biochem. 88, 135–141. [DOI] [PubMed] [Google Scholar]
  34. [34]Middleton, P., Dickson, D. P. E., Johnson, C. E. & Rush, J. D. (1980) Eur. J. Biochem. 104, 289–296. [DOI] [PubMed] [Google Scholar]
  35. [35]Moura, J. J. G., Moura, I., Kent, T. A., Lipscomb, J. D., Huynh, B. H., LeGall, J., Xavier, A. V. & Münck, E. (1982) J. Biol. Chem. 257, 6259–6267. [PubMed] [Google Scholar]
  36. [36]Hu, Z., Jollie, D., Burgess, B. K., Stephens, P. J. & Münck, E. (1994) Biochemistry 33, 14475–14485. [DOI] [PubMed] [Google Scholar]
  37. [37]Yoo, S. J., Angove, H. C., Burgess, B. K., Hendrich, M. P. & Münck, E. (1999) J. Am. Chem. Soc. 121, 2534–2545. [Google Scholar]
  38. [38]Pereira, A. S., Tavares, P., Moura, I., Moura, J. J. G. & Huynh, B. H. (2001) J. Am. Chem. Soc. 123, 2771–2782. [DOI] [PubMed] [Google Scholar]
  39. [39]Achim, C., Bominaar, E. L., Meyer, J., Peterson, J. & Münck, E. (1999) J. Am. Chem. Soc. 121, 3704–3714. [Google Scholar]
  40. [40]Pufahl, R. A., Singer, C. P., Pearisco, K. L., Lin, S.-J., Schmidt, P. J., Fahrni, C. J., Cizewski-Culotta, V., Penner-Hahn, J. E. & O'Halloran, T. V. (1997) Science 278, 853–856. [DOI] [PubMed] [Google Scholar]
  41. [41]Popescu, C. V., Bates, D. M., Beinert, H., Munck, E. & Kiley, P. J. (1998) Proc. Natl. Acad. Sci. USA 95, 13431–13435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. [42]Wu, G., Mansy, S. S., Wu, S.-P., Surerus, K. K., Foster, M. W. & Cowan, J. A. (2002) Biochemistry 41, 5024–5032. [DOI] [PubMed] [Google Scholar]
  43. [43]Nuth, M., Yoon, T. & Cowan, J. A. (2002) J. Am. Chem. Soc. 124, 8774–8775. [DOI] [PubMed] [Google Scholar]
  44. [44]Mansy, S. S., Wu, G., Surerus, K. K. & Cowan, J. A. (2002) J. Biol. Chem. 277, 21397–21404. [DOI] [PubMed] [Google Scholar]
  45. [45]Mihara, H. & Esaki, N. (2002) Appl. Microbiol. Biotechnol. 60, 12–23. [DOI] [PubMed] [Google Scholar]
  46. [46]Strain, J., Lorenz, C. R., Bode, J., Garland, S., Smolen, G. A., Ta, D. T., Vickery, L. E. & Culotta, V. C. (1998) J. Biol. Chem. 273, 31138–31144. [DOI] [PubMed] [Google Scholar]
  47. [47]Knight, S. A., Sepuri, N. B., Pain, D. & Dancis, A. (1998) J. Biol. Chem. 273, 18389–18393. [DOI] [PubMed] [Google Scholar]
  48. [48]Lange, H., Kaut, A., Kispal, G. & Lill, R. (2000) Proc. Natl. Acad. Sci. USA 97, 1050–1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. [49]Voisine, C., Cheng, Y. C., Ohlson, M., Schilke, B., Hoff, K., Beinert, H., Marszalek, J. & Craig, E. A. (2001) Proc. Natl. Acad. Sci. USA 98, 1483–1488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. [50]Rouault, T. A. & Kalusner, R. D. (1996) Trends Biochem. Sci. 21, 174–177. [PubMed] [Google Scholar]
  51. [51]Kispal, G., Csere, P., Prohl, C. & Lill, R. (1999) EMBO J. 18, 3981–3989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. [52]Lill, R., Diekert, K., Kaut, A., Lange, H., Pelzer, W., Prohl, C. & Kispal, G. (1999) Biol. Chem. 380, 1157–1166. [DOI] [PubMed] [Google Scholar]
  53. [53]Van Ho, A., Ward, D. M. & Kaplan, J. (2002) Annu. Rev. Microbiol. 56, 237–261. [DOI] [PubMed] [Google Scholar]
  54. [54]Nakai, Y., Nakai, M., Hayashi, H. & Kagamiyama, H. (2001) J. Biol. Chem. 276, 8314–8320. [DOI] [PubMed] [Google Scholar]

Cited by

Sorted by citations

Related

Authors
Tong, W.-H., Jameson, G. N. L., Huynh, B. H., & Rouault, T. A.
Affiliations
Emory University (US)
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2003-07-28
Volume
100
Issue
17
Pages
9762-9767
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.1732541100
PMC
PMC187839
OpenAlex
W2088723535
Cited by
194
References
54
Citations by year
2025: 3 · 2024: 4 · 2023: 3 · 2022: 5 · 2021: 7 · 2020: 13 · 2019: 7 · 2018: 16 · 2017: 17 · 2016: 15 · 2015: 4 · 2014: 6 · 2013: 12 · 2012: 4
FWCI
10.83
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • National Institute of General Medical Sciences (GM-47295, R01GM047295)
  • National Institute of Child Health and Human Development
Topics
  • Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
  • Physical Sciences › Chemistry › Inorganic Chemistry › Inorganic Chemistry and Materials
Keywords
iron-sulfur cluster assembly, scaffold proteins, [4Fe-4S] cluster, alternative splicing, subcellular compartmentalization, mitochondria, cytosol, nucleus, IscU, Isca, IscS, 57Fe Mössbauer spectroscopy
MeSH terms
Amino Acid Sequence, Cell Compartmentation, Humans, Iron-Sulfur Proteins, Molecular Sequence Data, Recombinant Proteins, RNA, Messenger, Spectrophotometry, Subcellular Fractions, Spectroscopy, Mossbauer, Molecular Structure, Alternative Splicing, Protein Isoforms, In Vitro Techniques

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review