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. 1992 May;12(5):1915–1923. doi: 10.1128/mcb.12.5.1915

A high-mobility-group protein and its cDNAs from Drosophila melanogaster.

C R Wagner 1, K Hamana 1, S C Elgin 1
PMCID: PMC364345  PMID: 1373803

Abstract

We have identified, purified, and characterized a high-mobility-group (HMG) protein and its cDNAs from Drosophila melanogaster. This protein, HMG D, shares most of the characteristics of vertebrate HMG proteins; it is extractable from nuclei with 0.35 M NaCl, is soluble in 5% perchloric acid, is relatively small (molecular weight of 12,000), has both a high basic (24%) and high acidic (24%) amino acid content, and is a DNA-binding protein. HMG D exhibits characteristics of both the vertebrate HMG 1 and 2 class and the HMG 14 and 17 class of proteins. Its amino acid sequence is similar (36% amino acid identity) to that of HMG1, while its size and selective extraction with ethidium bromide are similar to properties of the HMG 14 and 17 class of proteins. HMG D is encoded by a single-copy gene that maps to 57F8-11 on the right arm of chromosome 2. Two transcripts are observed during embryogenesis; the protein is relatively stable throughout development. By the biochemical criteria of size, solubility, and amino acid content, HMG D appears to be the major HMG protein of D. melanogaster.

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Selected References

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

  1. Abercrombie B. D., Kneale G. G., Crane-Robinson C., Bradbury E. M., Goodwin G. H., Walker J. M., Johns E. W. Studies on the conformational properties of the high-mobility-group chromosomal protein HMG 17 and its interaction with DNA. Eur J Biochem. 1978 Mar;84(1):173–177. doi: 10.1111/j.1432-1033.1978.tb12154.x. [DOI] [PubMed] [Google Scholar]
  2. Ashley C. T., Pendleton C. G., Jennings W. W., Saxena A., Glover C. V. Isolation and sequencing of cDNA clones encoding Drosophila chromosomal protein D1. A repeating motif in proteins which recognize at DNA. J Biol Chem. 1989 May 15;264(14):8394–8401. [PubMed] [Google Scholar]
  3. Bassuk J. A., Mayfield J. E. Major high mobility group like proteins of Drosophila melanogaster embryonic nuclei. Biochemistry. 1982 Mar 2;21(5):1024–1027. doi: 10.1021/bi00534a030. [DOI] [PubMed] [Google Scholar]
  4. Bianchi M. E., Beltrame M., Paonessa G. Specific recognition of cruciform DNA by nuclear protein HMG1. Science. 1989 Feb 24;243(4894 Pt 1):1056–1059. doi: 10.1126/science.2922595. [DOI] [PubMed] [Google Scholar]
  5. Brown N. H., Kafatos F. C. Functional cDNA libraries from Drosophila embryos. J Mol Biol. 1988 Sep 20;203(2):425–437. doi: 10.1016/0022-2836(88)90010-1. [DOI] [PubMed] [Google Scholar]
  6. Bustin M., Lehn D. A., Landsman D. Structural features of the HMG chromosomal proteins and their genes. Biochim Biophys Acta. 1990 Jul 30;1049(3):231–243. doi: 10.1016/0167-4781(90)90092-g. [DOI] [PubMed] [Google Scholar]
  7. Cary P. D., King D. S., Crane-Robinson C., Bradbury E. M., Rabbani A., Goodwin G. H., Johns E. W. Structural studies on two high-mobility-group proteins from calf thymus, HMG-14 and HMG-20 (ubiquitin), and their interaction with DNA. Eur J Biochem. 1980 Dec;112(3):577–580. doi: 10.1111/j.1432-1033.1980.tb06123.x. [DOI] [PubMed] [Google Scholar]
  8. Cavener D. R. Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res. 1987 Feb 25;15(4):1353–1361. doi: 10.1093/nar/15.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Croston G. E., Kerrigan L. A., Lira L. M., Marshak D. R., Kadonaga J. T. Sequence-specific antirepression of histone H1-mediated inhibition of basal RNA polymerase II transcription. Science. 1991 Feb 8;251(4994):643–649. doi: 10.1126/science.1899487. [DOI] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  13. Gilmour D. S., Dietz T. J., Elgin S. C. TATA box-dependent protein-DNA interactions are detected on heat shock and histone gene promoters in nuclear extracts derived from Drosophila melanogaster embryos. Mol Cell Biol. 1988 Aug;8(8):3204–3214. doi: 10.1128/mcb.8.8.3204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Goodwin G. H., Nicolas R. H., Johns E. W. An improved large scale fractionation of high mobility group non-histone chromatin proteins. Biochim Biophys Acta. 1975 Oct 20;405(2):280–291. doi: 10.1016/0005-2795(75)90094-x. [DOI] [PubMed] [Google Scholar]
  15. Goodwin G. H., Sanders C., Johns E. W. A new group of chromatin-associated proteins with a high content of acidic and basic amino acids. Eur J Biochem. 1973 Sep 21;38(1):14–19. doi: 10.1111/j.1432-1033.1973.tb03026.x. [DOI] [PubMed] [Google Scholar]
  16. Haggren W., Kolodrubetz D. The Saccharomyces cerevisiae ACP2 gene encodes an essential HMG1-like protein. Mol Cell Biol. 1988 Mar;8(3):1282–1289. doi: 10.1128/mcb.8.3.1282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hamana K., Iwai K. High mobility group nonhistone chromosomal proteins also exist in Tetrahymena. J Biochem. 1979 Sep;86(3):789–794. doi: 10.1093/oxfordjournals.jbchem.a132586. [DOI] [PubMed] [Google Scholar]
  18. Harrison S. D., Travers A. A. Identification of the binding sites for potential regulatory proteins in the upstream enhancer element of the Drosophila fushi tarazu gene. Nucleic Acids Res. 1988 Dec 23;16(24):11403–11416. doi: 10.1093/nar/16.24.11403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
  20. Hunkapiller M. W., Lujan E., Ostrander F., Hood L. E. Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. Methods Enzymol. 1983;91:227–236. doi: 10.1016/s0076-6879(83)91019-4. [DOI] [PubMed] [Google Scholar]
  21. Isackson P. J., Fishback J. L., Bidney D. L., Reeck G. R. Preferential affinity of high molecular weight high mobility group non-histone chromatin proteins for single-stranded DNA. J Biol Chem. 1979 Jul 10;254(13):5569–5572. [PubMed] [Google Scholar]
  22. Jantzen H. M., Admon A., Bell S. P., Tjian R. Nucleolar transcription factor hUBF contains a DNA-binding motif with homology to HMG proteins. Nature. 1990 Apr 26;344(6269):830–836. doi: 10.1038/344830a0. [DOI] [PubMed] [Google Scholar]
  23. Johns E. W. Studies on histones. 7. Preparative methods for histone fractions from calf thymus. Biochem J. 1964 Jul;92(1):55–59. doi: 10.1042/bj0920055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Johnson K. R., Lehn D. A., Elton T. S., Barr P. J., Reeves R. Complete murine cDNA sequence, genomic structure, and tissue expression of the high mobility group protein HMG-I(Y). J Biol Chem. 1988 Dec 5;263(34):18338–18342. [PubMed] [Google Scholar]
  25. Kolodrubetz D., Burgum A. Duplicated NHP6 genes of Saccharomyces cerevisiae encode proteins homologous to bovine high mobility group protein 1. J Biol Chem. 1990 Feb 25;265(6):3234–3239. [PubMed] [Google Scholar]
  26. Kruger W., Herskowitz I. A negative regulator of HO transcription, SIN1 (SPT2), is a nonspecific DNA-binding protein related to HMG1. Mol Cell Biol. 1991 Aug;11(8):4135–4146. doi: 10.1128/mcb.11.8.4135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Landsman D., Bustin M. Chicken non-histone chromosomal protein HMG-17 cDNA sequence. Nucleic Acids Res. 1987 Aug 25;15(16):6750–6750. doi: 10.1093/nar/15.16.6750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Levy-Wilson B., Denker M. S., Ito E. Isolation, characterization, and postsynthetic modifications of tetrahymena high mobility group proteins. Biochemistry. 1983 Mar 29;22(7):1715–1721. doi: 10.1021/bi00276a030. [DOI] [PubMed] [Google Scholar]
  29. Mardian J. K., Paton A. E., Bunick G. J., Olins D. E. Nucleosome cores have two specific binding sites for nonhistone chromosomal proteins HMG 14 and HMG 17. Science. 1980 Sep 26;209(4464):1534–1536. doi: 10.1126/science.7433974. [DOI] [PubMed] [Google Scholar]
  30. Mathew C. G., Goodwin G. H., Johns E. W. Studies on the association of the high mobility group non-histone chromatin proteins with isolated nucleosomes. Nucleic Acids Res. 1979 Jan;6(1):167–179. doi: 10.1093/nar/6.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  32. Pikaard C. S., McStay B., Schultz M. C., Bell S. P., Reeder R. H. The Xenopus ribosomal gene enhancers bind an essential polymerase I transcription factor, xUBF. Genes Dev. 1989 Nov;3(11):1779–1788. doi: 10.1101/gad.3.11.1779. [DOI] [PubMed] [Google Scholar]
  33. Rodriguez Alfageme C., Rudkin G. T., Cohen L. H. Isolation, properties and cellular distribution of D1, a chromosomal protein of Drosophila. Chromosoma. 1980;78(1):1–31. doi: 10.1007/BF00291907. [DOI] [PubMed] [Google Scholar]
  34. Roeder G. S., Beard C., Smith M., Keranen S. Isolation and characterization of the SPT2 gene, a negative regulator of Ty-controlled yeast gene expression. Mol Cell Biol. 1985 Jul;5(7):1543–1553. doi: 10.1128/mcb.5.7.1543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sandeen G., Wood W. I., Felsenfeld G. The interaction of high mobility proteins HMG14 and 17 with nucleosomes. Nucleic Acids Res. 1980 Sep 11;8(17):3757–3778. doi: 10.1093/nar/8.17.3757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sanders C. A method for the fractionation of the high-mobility-group non-histome chromosomal proteins. Biochem Biophys Res Commun. 1977 Oct 10;78(3):1034–1042. doi: 10.1016/0006-291x(77)90525-3. [DOI] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schröter H., Maier G., Ponstingl H., Nordheim A. DNA intercalators induce specific release of HMG 14, HMG 17 and other DNA-binding proteins from chicken erythrocyte chromatin. EMBO J. 1985 Dec 30;4(13B):3867–3872. doi: 10.1002/j.1460-2075.1985.tb04159.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schulman I. G., Cook R. G., Richman R., Allis C. D. Tetrahymena contain two distinct and unusual high mobility group (HMG)-like proteins. J Cell Biol. 1987 Jun;104(6):1485–1494. doi: 10.1083/jcb.104.6.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schulman I. G., Wang T., Wu M., Bowen J., Cook R. G., Gorovsky M. A., Allis C. D. Macronuclei and micronuclei in Tetrahymena thermophila contain high-mobility-group-like chromosomal proteins containing a highly conserved eleven-amino-acid putative DNA-binding sequence. Mol Cell Biol. 1991 Jan;11(1):166–174. doi: 10.1128/mcb.11.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shick V. V., Belyavsky A. V., Mirzabekov A. D. Primary organization of nucleosomes. Interaction of non-histone high mobility group proteins 14 and 17 with nucleosomes, as revealed by DNA-protein crosslinking and immunoaffinity isolation. J Mol Biol. 1985 Sep 20;185(2):329–339. doi: 10.1016/0022-2836(85)90407-3. [DOI] [PubMed] [Google Scholar]
  42. Solomon M. J., Strauss F., Varshavsky A. A mammalian high mobility group protein recognizes any stretch of six A.T base pairs in duplex DNA. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1276–1280. doi: 10.1073/pnas.83.5.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vaslet C. A., O'Connell P., Izquierdo M., Rosbash M. Isolation and mapping of a cloned ribosomal protein gene of Drosophila melanogaster. Nature. 1980 Jun 26;285(5767):674–676. doi: 10.1038/285674a0. [DOI] [PubMed] [Google Scholar]
  44. Waterman M. L., Fischer W. H., Jones K. A. A thymus-specific member of the HMG protein family regulates the human T cell receptor C alpha enhancer. Genes Dev. 1991 Apr;5(4):656–669. doi: 10.1101/gad.5.4.656. [DOI] [PubMed] [Google Scholar]
  45. Weisbrod S., Weintraub H. Isolation of a subclass of nuclear proteins responsible for conferring a DNase I-sensitive structure on globin chromatin. Proc Natl Acad Sci U S A. 1979 Feb;76(2):630–634. doi: 10.1073/pnas.76.2.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wustmann G., Szidonya J., Taubert H., Reuter G. The genetics of position-effect variegation modifying loci in Drosophila melanogaster. Mol Gen Genet. 1989 Jun;217(2-3):520–527. doi: 10.1007/BF02464926. [DOI] [PubMed] [Google Scholar]

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