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. 1994 Jan 4;91(1):316–320. doi: 10.1073/pnas.91.1.316

Cysteine-rich LIM domains of LIM-homeodomain and LIM-only proteins contain zinc but not iron.

V E Archer 1, J Breton 1, I Sanchez-Garcia 1, H Osada 1, A Forster 1, A J Thomson 1, T H Rabbitts 1
PMCID: PMC42938  PMID: 7904068

Abstract

The structure of LIM domains has major implications for transcription because proteins such as Is1-1 contain two LIM domains associated with a homeodomain, and RBTN1/Ttg-1 and RBTN2/Ttg-2 contain two LIM domains but no homeodomain. Conserved cysteine and histidine residues in the LIM domains suggest a metal-binding role. RBTN and Is1-1 LIM proteins have been made in Escherichia coli and insect cell expression systems and their metal content has been determined using atomic absorption spectroscopy and electron paramagnetic resonance spectroscopy. LIM proteins expressed in soluble form contain zinc atoms, whereas bacterial inclusion bodies invariably also have Fe-S clusters. The latter are identified as linear [Fe3S4]+ clusters and appear to result from incorrect metal coordination by E. coli. These studies show that RBTN1, RBTN2, and Is1-1 are metalloproteins that contain zinc but not iron and, therefore, that the LIM domain represents a zinc-binding domain.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blackwood E. M., Eisenman R. N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science. 1991 Mar 8;251(4998):1211–1217. doi: 10.1126/science.2006410. [DOI] [PubMed] [Google Scholar]
  2. Boehm T., Baer R., Lavenir I., Forster A., Waters J. J., Nacheva E., Rabbitts T. H. The mechanism of chromosomal translocation t(11;14) involving the T-cell receptor C delta locus on human chromosome 14q11 and a transcribed region of chromosome 11p15. EMBO J. 1988 Feb;7(2):385–394. doi: 10.1002/j.1460-2075.1988.tb02825.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boehm T., Foroni L., Kaneko Y., Perutz M. F., Rabbitts T. H. The rhombotin family of cysteine-rich LIM-domain oncogenes: distinct members are involved in T-cell translocations to human chromosomes 11p15 and 11p13. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4367–4371. doi: 10.1073/pnas.88.10.4367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boehm T., Greenberg J. M., Buluwela L., Lavenir I., Forster A., Rabbitts T. H. An unusual structure of a putative T cell oncogene which allows production of similar proteins from distinct mRNAs. EMBO J. 1990 Mar;9(3):857–868. doi: 10.1002/j.1460-2075.1990.tb08183.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bourdineaud J. P., Howard S. P., Pages J. M., Bernadac A., Leroy G., Bruschi M., Lazdunski C. Cytoplasmic and periplasmic expression of a synthetic gene for ferredoxin in Escherichia coli. Biochimie. 1990 Jun-Jul;72(6-7):407–415. doi: 10.1016/0300-9084(90)90065-o. [DOI] [PubMed] [Google Scholar]
  6. Crawford A. W., Michelsen J. W., Beckerle M. C. An interaction between zyxin and alpha-actinin. J Cell Biol. 1992 Mar;116(6):1381–1393. doi: 10.1083/jcb.116.6.1381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Freyd G., Kim S. K., Horvitz H. R. Novel cysteine-rich motif and homeodomain in the product of the Caenorhabditis elegans cell lineage gene lin-11. Nature. 1990 Apr 26;344(6269):876–879. doi: 10.1038/344876a0. [DOI] [PubMed] [Google Scholar]
  8. Guan K. L., Dixon J. E. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. doi: 10.1016/0003-2697(91)90534-z. [DOI] [PubMed] [Google Scholar]
  9. Harrison S. D., Travers A. A. The tramtrack gene encodes a Drosophila finger protein that interacts with the ftz transcriptional regulatory region and shows a novel embryonic expression pattern. EMBO J. 1990 Jan;9(1):207–216. doi: 10.1002/j.1460-2075.1990.tb08097.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hempe J. M., Cousins R. J. Cysteine-rich intestinal protein binds zinc during transmucosal zinc transport. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9671–9674. doi: 10.1073/pnas.88.21.9671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Karlsson O., Thor S., Norberg T., Ohlsson H., Edlund T. Insulin gene enhancer binding protein Isl-1 is a member of a novel class of proteins containing both a homeo- and a Cys-His domain. Nature. 1990 Apr 26;344(6269):879–882. doi: 10.1038/344879a0. [DOI] [PubMed] [Google Scholar]
  12. Kennedy M. C., Kent T. A., Emptage M., Merkle H., Beinert H., Münck E. Evidence for the formation of a linear [3Fe-4S] cluster in partially unfolded aconitase. J Biol Chem. 1984 Dec 10;259(23):14463–14471. [PubMed] [Google Scholar]
  13. Li P. M., Reichert J., Freyd G., Horvitz H. R., Walsh C. T. The LIM region of a presumptive Caenorhabditis elegans transcription factor is an iron-sulfur- and zinc-containing metallodomain. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9210–9213. doi: 10.1073/pnas.88.20.9210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Liebhaber S. A., Emery J. G., Urbanek M., Wang X. K., Cooke N. E. Characterization of a human cDNA encoding a widely expressed and highly conserved cysteine-rich protein with an unusual zinc-finger motif. Nucleic Acids Res. 1990 Jul 11;18(13):3871–3879. doi: 10.1093/nar/18.13.3871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McGuire E. A., Hockett R. D., Pollock K. M., Bartholdi M. F., O'Brien S. J., Korsmeyer S. J. The t(11;14)(p15;q11) in a T-cell acute lymphoblastic leukemia cell line activates multiple transcripts, including Ttg-1, a gene encoding a potential zinc finger protein. Mol Cell Biol. 1989 May;9(5):2124–2132. doi: 10.1128/mcb.9.5.2124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Michelsen J. W., Schmeichel K. L., Beckerle M. C., Winge D. R. The LIM motif defines a specific zinc-binding protein domain. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4404–4408. doi: 10.1073/pnas.90.10.4404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nagai K., Thøgersen H. C. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli. Methods Enzymol. 1987;153:461–481. doi: 10.1016/0076-6879(87)53072-5. [DOI] [PubMed] [Google Scholar]
  18. Omichinski J. G., Clore G. M., Schaad O., Felsenfeld G., Trainor C., Appella E., Stahl S. J., Gronenborn A. M. NMR structure of a specific DNA complex of Zn-containing DNA binding domain of GATA-1. Science. 1993 Jul 23;261(5120):438–446. doi: 10.1126/science.8332909. [DOI] [PubMed] [Google Scholar]
  19. Omichinski J. G., Trainor C., Evans T., Gronenborn A. M., Clore G. M., Felsenfeld G. A small single-"finger" peptide from the erythroid transcription factor GATA-1 binds specifically to DNA as a zinc or iron complex. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1676–1680. doi: 10.1073/pnas.90.5.1676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rabbitts T. H., Boehm T. LIM domains. Nature. 1990 Aug 2;346(6283):418–418. doi: 10.1038/346418a0. [DOI] [PubMed] [Google Scholar]
  21. Royer-Pokora B., Loos U., Ludwig W. D. TTG-2, a new gene encoding a cysteine-rich protein with the LIM motif, is overexpressed in acute T-cell leukaemia with the t(11;14)(p13;q11). Oncogene. 1991 Oct;6(10):1887–1893. [PubMed] [Google Scholar]
  22. Sadler I., Crawford A. W., Michelsen J. W., Beckerle M. C. Zyxin and cCRP: two interactive LIM domain proteins associated with the cytoskeleton. J Cell Biol. 1992 Dec;119(6):1573–1587. doi: 10.1083/jcb.119.6.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schwabe J. W., Neuhaus D., Rhodes D. Solution structure of the DNA-binding domain of the oestrogen receptor. Nature. 1990 Nov 29;348(6300):458–461. doi: 10.1038/348458a0. [DOI] [PubMed] [Google Scholar]
  24. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  25. Sánchez-García I., Osada H., Forster A., Rabbitts T. H. The cysteine-rich LIM domains inhibit DNA binding by the associated homeodomain in Isl-1. EMBO J. 1993 Nov;12(11):4243–4250. doi: 10.1002/j.1460-2075.1993.tb06108.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thomson A. J., Cheesman M. R., George S. J. Variable-temperature magnetic circular dichroism. Methods Enzymol. 1993;226:199–232. doi: 10.1016/0076-6879(93)26011-w. [DOI] [PubMed] [Google Scholar]
  27. Thomson A. J. Iron-sulphur proteins: crosslinked by a cluster. Curr Biol. 1993 Mar;3(3):173–174. doi: 10.1016/0960-9822(93)90264-o. [DOI] [PubMed] [Google Scholar]
  28. Way J. C., Chalfie M. mec-3, a homeobox-containing gene that specifies differentiation of the touch receptor neurons in C. elegans. Cell. 1988 Jul 1;54(1):5–16. doi: 10.1016/0092-8674(88)90174-2. [DOI] [PubMed] [Google Scholar]

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