Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1998 Apr 1;26(7):1605–1612. doi: 10.1093/nar/26.7.1605

A conserved structural element in horse and mouse IGF2 genes binds a methylation sensitive factor.

K Otte 1, D Choudhury 1, M Charalambous 1, W Engström 1, B Rozell 1
PMCID: PMC147450  PMID: 9512529

Abstract

The equine IGF2 gene has been cloned and characterised. It spans a 9 kb region, which is substantially less than the corresponding human gene. Three coding exons and three untranslated leader exons, all highly homologous to those in other species, were identified. Downstream of the polyadenylation site in exon 6, a dinucleotide repeat sequence was identified. Three putative promoters (P1-P3) were localised in the 5' region of the gene. RNase protection analysis revealed two active promoters in fetal tissues, P2 and P3, whereas P3 was the only promoter active in adult tissues. This represents a transcriptional pattern different from that in humans or rodents. A novel structural element, an inverted repeat, is predicted in the 3' region of the IGF2 gene. This repeat is conserved between species and located in a region which is differentially methylated in the human and mouse genes and might therefore be involved in the imprinting mechanism. The inverted repeat acquires a stem-loop structure in vitro with a hybrid A/B-DNA conformation in the stem area. Both in horse and mouse, a methylation-sensitive protein binds this structure with a strong requirement for the loop area. Furthermore, the protein might be developmentally regulated.

Full Text

The Full Text of this article is available as a PDF (257.3 KB).

Selected References

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

  1. Banerjee S., Smallwood A. A chromatin model of IGF2/H19 imprinting. Nat Genet. 1995 Nov;11(3):237–238. doi: 10.1038/ng1195-237. [DOI] [PubMed] [Google Scholar]
  2. Blundell T. L., Humbel R. E. Hormone families: pancreatic hormones and homologous growth factors. Nature. 1980 Oct 30;287(5785):781–787. doi: 10.1038/287781a0. [DOI] [PubMed] [Google Scholar]
  3. Brandeis M., Kafri T., Ariel M., Chaillet J. R., McCarrey J., Razin A., Cedar H. The ontogeny of allele-specific methylation associated with imprinted genes in the mouse. EMBO J. 1993 Sep;12(9):3669–3677. doi: 10.1002/j.1460-2075.1993.tb06041.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Christiansen J., Kofod M., Nielsen F. C. A guanosine quadruplex and two stable hairpins flank a major cleavage site in insulin-like growth factor II mRNA. Nucleic Acids Res. 1994 Dec 25;22(25):5709–5716. doi: 10.1093/nar/22.25.5709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Daughaday W. H., Rotwein P. Insulin-like growth factors I and II. Peptide, messenger ribonucleic acid and gene structures, serum, and tissue concentrations. Endocr Rev. 1989 Feb;10(1):68–91. doi: 10.1210/edrv-10-1-68. [DOI] [PubMed] [Google Scholar]
  6. De Moor C. H., Jansen M., Sussenbach J. S., Van den Brande J. L. Differential polysomal localization of human insulin-like-growth-factor-2 mRNAs in cell lines and foetal liver. Eur J Biochem. 1994 Jun 15;222(3):1017–1024. doi: 10.1111/j.1432-1033.1994.tb18953.x. [DOI] [PubMed] [Google Scholar]
  7. DeChiara T. M., Efstratiadis A., Robertson E. J. A growth-deficiency phenotype in heterozygous mice carrying an insulin-like growth factor II gene disrupted by targeting. Nature. 1990 May 3;345(6270):78–80. doi: 10.1038/345078a0. [DOI] [PubMed] [Google Scholar]
  8. DeChiara T. M., Robertson E. J., Efstratiadis A. Parental imprinting of the mouse insulin-like growth factor II gene. Cell. 1991 Feb 22;64(4):849–859. doi: 10.1016/0092-8674(91)90513-x. [DOI] [PubMed] [Google Scholar]
  9. DeChiara T. M., Robertson E. J., Efstratiadis A. Parental imprinting of the mouse insulin-like growth factor II gene. Cell. 1991 Feb 22;64(4):849–859. doi: 10.1016/0092-8674(91)90513-x. [DOI] [PubMed] [Google Scholar]
  10. Feil R., Walter J., Allen N. D., Reik W. Developmental control of allelic methylation in the imprinted mouse Igf2 and H19 genes. Development. 1994 Oct;120(10):2933–2943. doi: 10.1242/dev.120.10.2933. [DOI] [PubMed] [Google Scholar]
  11. Frunzio R., Chiariotti L., Brown A. L., Graham D. E., Rechler M. M., Bruni C. B. Structure and expression of the rat insulin-like growth factor II (rIGF-II) gene. rIGF-II RNAs are transcribed from two promoters. J Biol Chem. 1986 Dec 25;261(36):17138–17149. [PubMed] [Google Scholar]
  12. Giannoukakis N., Deal C., Paquette J., Goodyer C. G., Polychronakos C. Parental genomic imprinting of the human IGF2 gene. Nat Genet. 1993 May;4(1):98–101. doi: 10.1038/ng0593-98. [DOI] [PubMed] [Google Scholar]
  13. Grabczyk E., Fishman M. C. A long purine-pyrimidine homopolymer acts as a transcriptional diode. J Biol Chem. 1995 Jan 27;270(4):1791–1797. doi: 10.1074/jbc.270.4.1791. [DOI] [PubMed] [Google Scholar]
  14. Holthuizen P., van der Lee F. M., Ikejiri K., Yamamoto M., Sussenbach J. S. Identification and initial characterization of a fourth leader exon and promoter of the human IGF-II gene. Biochim Biophys Acta. 1990 Nov 30;1087(3):341–343. doi: 10.1016/0167-4781(90)90010-y. [DOI] [PubMed] [Google Scholar]
  15. Horwitz M. S., Loeb L. A. An E. coli promoter that regulates transcription by DNA superhelix-induced cruciform extrusion. Science. 1988 Aug 5;241(4866):703–705. doi: 10.1126/science.2456617. [DOI] [PubMed] [Google Scholar]
  16. Ikejiri K., Furuichi M., Ueno T., Matsuguchi T., Takahashi K., Endo H., Yamamoto M. The presence and active transcription of three independent leader exons in the mouse insulin-like growth factor II gene. Biochim Biophys Acta. 1991 May 2;1089(1):77–82. doi: 10.1016/0167-4781(91)90087-3. [DOI] [PubMed] [Google Scholar]
  17. Ikejiri K., Ueno T., Matsuguchi T., Takahashi K., Endo H., Yamamoto M. The primary structure of the rat insulin-like growth factor II gene region. Biochim Biophys Acta. 1990 Jul 30;1049(3):350–353. doi: 10.1016/0167-4781(90)90110-n. [DOI] [PubMed] [Google Scholar]
  18. Ikejiri K., Wasada T., Haruki K., Hizuka N., Hirata Y., Yamamoto M. Identification of a novel transcription unit in the human insulin-like growth factor-II gene. Biochem J. 1991 Dec 1;280(Pt 2):439–444. doi: 10.1042/bj2800439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jin I. H., Sinha G., Yballe C., Vu T. H., Hoffman A. R. The human insulin-like growth factor-II promoter P1 is not restricted to liver: evidence for expression of P1 in other tissues and for a homologous promoter in baboon liver. Horm Metab Res. 1995 Oct;27(10):447–449. doi: 10.1055/s-2007-979999. [DOI] [PubMed] [Google Scholar]
  20. Kalb V. F., Jr, Bernlohr R. W. A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977 Oct;82(2):362–371. doi: 10.1016/0003-2697(77)90173-7. [DOI] [PubMed] [Google Scholar]
  21. Li E., Beard C., Jaenisch R. Role for DNA methylation in genomic imprinting. Nature. 1993 Nov 25;366(6453):362–365. doi: 10.1038/366362a0. [DOI] [PubMed] [Google Scholar]
  22. Li X., Cui H., Sandstedt B., Nordlinder H., Larsson E., Ekström T. J. Expression levels of the insulin-like growth factor-II gene (IGF2) in the human liver: developmental relationships of the four promoters. J Endocrinol. 1996 Apr;149(1):117–124. doi: 10.1677/joe.0.1490117. [DOI] [PubMed] [Google Scholar]
  23. Liu K., Miles H. T., Frazier J., Sasisekharan V. A novel DNA duplex. A parallel-stranded DNA helix with Hoogsteen base pairing. Biochemistry. 1993 Nov 9;32(44):11802–11809. doi: 10.1021/bi00095a008. [DOI] [PubMed] [Google Scholar]
  24. Manzini G., Xodo L. E., Gasparotto D., Quadrifoglio F., van der Marel G. A., van Boom J. H. Triple helix formation by oligopurine-oligopyrimidine DNA fragments. Electrophoretic and thermodynamic behavior. J Mol Biol. 1990 Jun 20;213(4):833–843. doi: 10.1016/S0022-2836(05)80267-0. [DOI] [PubMed] [Google Scholar]
  25. McMurray C. T., Wilson W. D., Douglass J. O. Hairpin formation within the enhancer region of the human enkephalin gene. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):666–670. doi: 10.1073/pnas.88.2.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moore T., Constancia M., Zubair M., Bailleul B., Feil R., Sasaki H., Reik W. Multiple imprinted sense and antisense transcripts, differential methylation and tandem repeats in a putative imprinting control region upstream of mouse Igf2. Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12509–12514. doi: 10.1073/pnas.94.23.12509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neumann B., Kubicka P., Barlow D. P. Characteristics of imprinted genes. Nat Genet. 1995 Jan;9(1):12–13. doi: 10.1038/ng0195-12. [DOI] [PubMed] [Google Scholar]
  28. Newell S., Ward A., Graham C. Discriminating translation of insulin-like growth factor-II (IGF-II) during mouse embryogenesis. Mol Reprod Dev. 1994 Nov;39(3):249–258. doi: 10.1002/mrd.1080390302. [DOI] [PubMed] [Google Scholar]
  29. Nielsen F. C., Gammeltoft S., Christiansen J. Translational discrimination of mRNAs coding for human insulin-like growth factor II. J Biol Chem. 1990 Aug 15;265(23):13431–13434. [PubMed] [Google Scholar]
  30. Nielsen F. C., Ostergaard L., Nielsen J., Christiansen J. Growth-dependent translation of IGF-II mRNA by a rapamycin-sensitive pathway. Nature. 1995 Sep 28;377(6547):358–362. doi: 10.1038/377358a0. [DOI] [PubMed] [Google Scholar]
  31. Ohlsen S. M., Lugenbeel K. A., Wong E. A. Characterization of the linked ovine insulin and insulin-like growth factor-II genes. DNA Cell Biol. 1994 Apr;13(4):377–388. doi: 10.1089/dna.1994.13.377. [DOI] [PubMed] [Google Scholar]
  32. Ohlsson R., Nyström A., Pfeifer-Ohlsson S., Töhönen V., Hedborg F., Schofield P., Flam F., Ekström T. J. IGF2 is parentally imprinted during human embryogenesis and in the Beckwith-Wiedemann syndrome. Nat Genet. 1993 May;4(1):94–97. doi: 10.1038/ng0593-94. [DOI] [PubMed] [Google Scholar]
  33. Otte K., Engström W. Insulin-like growth factor II in the horse: determination of a cDNA nucleotide sequence and expression in fetal and adult tissue. Gen Comp Endocrinol. 1994 Nov;96(2):270–275. doi: 10.1006/gcen.1994.1182. [DOI] [PubMed] [Google Scholar]
  34. Pearson C. E., Zorbas H., Price G. B., Zannis-Hadjopoulos M. Inverted repeats, stem-loops, and cruciforms: significance for initiation of DNA replication. J Cell Biochem. 1996 Oct;63(1):1–22. doi: 10.1002/(SICI)1097-4644(199610)63:1%3C1::AID-JCB1%3E3.0.CO;2-3. [DOI] [PubMed] [Google Scholar]
  35. Pedone P. V., Cosma M. P., Ungaro P., Colantuoni V., Bruni C. B., Zarrilli R., Riccio A. Parental imprinting of rat insulin-like growth factor II gene promoters is coordinately regulated. J Biol Chem. 1994 Sep 30;269(39):23970–23975. [PubMed] [Google Scholar]
  36. Riazance J. H., Baase W. A., Johnson W. C., Jr, Hall K., Cruz P., Tinoco I., Jr Evidence for Z-form RNA by vacuum UV circular dichroism. Nucleic Acids Res. 1985 Jul 11;13(13):4983–4989. doi: 10.1093/nar/13.13.4983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rinderknecht E., Humbel R. E. Primary structure of human insulin-like growth factor II. FEBS Lett. 1978 May 15;89(2):283–286. doi: 10.1016/0014-5793(78)80237-3. [DOI] [PubMed] [Google Scholar]
  38. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  39. Rotwein P., Hall L. J. Evolution of insulin-like growth factor II: characterization of the mouse IGF-II gene and identification of two pseudo-exons. DNA Cell Biol. 1990 Dec;9(10):725–735. doi: 10.1089/dna.1990.9.725. [DOI] [PubMed] [Google Scholar]
  40. Sasaki H., Jones P. A., Chaillet J. R., Ferguson-Smith A. C., Barton S. C., Reik W., Surani M. A. Parental imprinting: potentially active chromatin of the repressed maternal allele of the mouse insulin-like growth factor II (Igf2) gene. Genes Dev. 1992 Oct;6(10):1843–1856. doi: 10.1101/gad.6.10.1843. [DOI] [PubMed] [Google Scholar]
  41. Scheper W., Holthuizen P. E., Sussenbach J. S. The cis-acting elements involved in endonucleolytic cleavage of the 3' UTR of human IGF-II mRNAs bind a 50 kDa protein. Nucleic Acids Res. 1996 Mar 15;24(6):1000–1007. doi: 10.1093/nar/24.6.1000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schneid H., Seurin D., Vazquez M. P., Gourmelen M., Cabrol S., Le Bouc Y. Parental allele specific methylation of the human insulin-like growth factor II gene and Beckwith-Wiedemann syndrome. J Med Genet. 1993 May;30(5):353–362. doi: 10.1136/jmg.30.5.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Schofield P. N., Tate V. E. Regulation of human IGF-II transcription in fetal and adult tissues. Development. 1987 Dec;101(4):793–803. doi: 10.1242/dev.101.4.793. [DOI] [PubMed] [Google Scholar]
  44. Soares M. B., Turken A., Ishii D., Mills L., Episkopou V., Cotter S., Zeitlin S., Efstratiadis A. Rat insulin-like growth factor II gene. A single gene with two promoters expressing a multitranscript family. J Mol Biol. 1986 Dec 20;192(4):737–752. doi: 10.1016/0022-2836(86)90025-2. [DOI] [PubMed] [Google Scholar]
  45. Spiro C., Richards J. P., Chandrasekaran S., Brennan R. G., McMurray C. T. Secondary structure creates mismatched base pairs required for high-affinity binding of cAMP response element-binding protein to the human enkephalin enhancer. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4606–4610. doi: 10.1073/pnas.90.10.4606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Ueno T., Takahashi K., Matsuguchi T., Endo H., Yamamoto M. A new leader exon identified in the rat insulin-like growth factor II gene. Biochem Biophys Res Commun. 1987 Oct 14;148(1):344–349. doi: 10.1016/0006-291x(87)91116-8. [DOI] [PubMed] [Google Scholar]
  47. Ward A. Beck-Wiedemann syndrome and Wilms' tumour. Mol Hum Reprod. 1997 Feb;3(2):157–168. doi: 10.1093/molehr/3.2.157. [DOI] [PubMed] [Google Scholar]
  48. Zazopoulos E., Lalli E., Stocco D. M., Sassone-Corsi P. DNA binding and transcriptional repression by DAX-1 blocks steroidogenesis. Nature. 1997 Nov 20;390(6657):311–315. doi: 10.1038/36899. [DOI] [PubMed] [Google Scholar]
  49. de Pagter-Holthuizen P., Jansen M., van Schaik F. M., van der Kammen R., Oosterwijk C., Van den Brande J. L., Sussenbach J. S. The human insulin-like growth factor II gene contains two development-specific promoters. FEBS Lett. 1987 Apr 20;214(2):259–264. doi: 10.1016/0014-5793(87)80066-2. [DOI] [PubMed] [Google Scholar]
  50. de Pagter-Holthuizen P., Jansen M., van der Kammen R. A., van Schaik F. M., Sussenbach J. S. Differential expression of the human insulin-like growth factor II gene. Characterization of the IGF-II mRNAs and an mRNA encoding a putative IGF-II-associated protein. Biochim Biophys Acta. 1988 Sep 7;950(3):282–295. doi: 10.1016/0167-4781(88)90124-8. [DOI] [PubMed] [Google Scholar]
  51. van Dijk M. A., van Schaik F. M., Bootsma H. J., Holthuizen P., Sussenbach J. S. Initial characterization of the four promoters of the human insulin-like growth factor II gene. Mol Cell Endocrinol. 1991 Oct;81(1-3):81–94. doi: 10.1016/0303-7207(91)90207-9. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES