Identification of Grf1 on mouse chromosome 9 as an imprinted gene by RLGS–M

Abstract

Normal mammalian development requires a diploid combination of both haploid parental genomes1. Uniparental disomy for certain segments of specific chromosomes results in aberrant development or prenatal lethality2,3, indicating that the parental genomes have undergone modifications during gametogenesis. These modifications result in parent-of-origin specific expression for some genes, a phenomenon called genomic imprinting. Recent work with DNA methyltransferase deficient mice showed that differential methylation is the probable basis of the imprinted character of several genes4. Screening for endogenous imprinted loci using restriction landmark genomic scanning with methylation sensitive enzymes (RLGS-M) identified eight imprinted RLGS (Irlgs) candidate loci5,6. Molecular analysis of the genomic region of one of the loci (Irigs2) resulted in the discovery of the paternally imprinted U2afbp-rs gene within a previously identified imprinted region on mouse chromosome 11 (refs 5, 7). This paper describes the characterisation of a novel imprinted RLGS-M locus, Irlgs3, on mouse chromosome 9 (ref. 6). Within this locus we identified the Grf1 (also called Cdc25Mm) gene, which is homologous to the RAS-specific guanine nucleotide exchange factor gene, CDC25, in Saccharomyces cerevisiae. Grf1 is located about 30 kb downstream of the methylation imprinted site, identified by RLGS-M, and shows paternal allele specific expression in mouse brain, stomach and heart. Our results indicate that imprinting may have a role in regulating mitogenic signal transduction pathways during growth and development.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Softer, D. Differential imprinting and expression of maternal and paternal genomes. Ainu. Rev. Genet. 22, 127–146 (1988).

  2. 2

    Beechey, C.V. & Cattanach, B.M. Genetic imprinting map. Mouse Genome 91, 102–104 (1993).

  3. 3

    Cattanach, B.M. Parental origin effects in mice. J. Embtyol. Exp. Morph. 97, 137–150 (1986).

  4. 4

    Li, E., Beard, C. & Jaenisch, R. Role for DNA methylation in genomic imprinting. Nature 366, 362–365 (1993).

  5. 5

    Hayashizaki, Y. et al. Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method. Nature Genet 6, 33–40 (1994).

  6. 6

    Shibata, H. et al. Genetic mapping and systematic screening of mouse endogenously imprinted loci detected with restriction landmark genome scanning method (RLGS). Mamm. Genome 5, 797–800 (1994).

  7. 7

    Kalcheva, I. et al. Comparative mapping of the imprinted U2afbpL gene on mouse chromosome 11 and human chromosome 5. Cytogenet. Cell Genet. 68, 19–24 (1995).

  8. 8

    Shibata, H. et al. The use of restriction landmark genomic scanning to scan the mouse genome for endogenous loci with imprinted patterns of methylation. Eectrophoresis 16, 210–217 (1995).

  9. 9

    Cen, H., Papageorge, A.G., Zippel, R., Lowy, D.R. & Zhang, K. Isolation of multiple mouse cDNAs with coding homology to Saccharomyces cerevisiae CDC25: identification of a region related to Bcr, Vav, Dbl and CDC24 . EMBO J. 11, 4007–4015 (1992).

  10. 10

    Pierce, J.C. & Sternberg, N. Using the bacteriophage P1 system to clone high molecular weight (HMW) genomic DNA. Meth. Enzymol. 216, 549–574 (1992).

  11. 11

    Leighton, P.A., Ingram, R.S., Eggenschwiller, J., Efstratiadis, A. & Tilghman, S.M. Disruption of imprinting caused by deletion of the H19 gene region in mice. Nature 375, 34–39 (1995).

  12. 12

    Razin, A. & Cedar, H. DNA Methylation and gene expression. Microbiol. Rev. 55, 451–458 (1991).

  13. 13

    Stoeger, R. et al. Maternal-specific methylation of the imprinted mouse Igf2r locus identifies the expressed locus as carrying the imprinting signal. Cell 73, 61–71 (1993).

  14. 14

    Feil, R., Walter, J., Allen, N.D. & Reik, W. Developmental control of allelic methylation in the imprinted mouse Igf2 and H19 genes. Development 120, 2933–2943 (1994).

  15. 15

    Gariboldi, M. et al. Genetic mapping of the mouse CDC25Mm gene, a ras-specific guanine nucleotide-releasing factor, to chromosome 9. Genomics 21, 451–453 (1994).

  16. 16

    Beechey, C. & Cattanach, B.M. Genetic imprinting map. Mouse Genome 94, 96–99 (1996).

  17. 17

    Giddings, S.J., King, C.D., Harman, K.W., Flood, J.F. & Carnaghi, L.R. Allele specific inactivation of insulin 1 and 2, in the mouse yolk sac, indicates imprinting. Nature Genet. 6, 310–313 (1994).

  18. 18

    Cen, H., Papageorge, A.G., Vass, W.C., Zhang, K. & Lowy, D.R. Regulated and constitutive activity by CDC25Mm (GRF), Ras-specific exchange factor. Mol. Cell. Biol. 13, 7718–7724 (1993).

  19. 19

    Villar, A. & Pedersen, R.A. Parental imprinting of the Mas protooncogene in mouse. Nature Genet. 8, 373–379 (1994).

  20. 20

    Hatada, I. & Mukai, T. Genomic imprinting of p57KIP2, a cyclin-dependent kinase inhibitor, in mouse. Nature Genet. 11, 204–206 (1995).

  21. 21

    Lau, M.M.H. et al. Loss of the imprinted lGF2/cation-independent mannose 6-phosphate receptor results in fetal overgrowth and perinatal lethality. Genes Dev. 8, 2953–2963 (1994).

  22. 22

    DeChiara, T.M., Robertson, E.J. & Efstratiadis, A. Parental imprinting of the mouse insulin-like growth factor II gene. Cell 64, 849–859 (1991).

  23. 23

    Wang, Z.-Q., Fung, M.R., Barlow, D.P. & Wagner, E.F. Regulation of embryonic growth and lysosomal targeting by the imprinted lgf/Mpr gene. Nature 372, 464–467 (1994).

  24. 24

    Hirotsune, S. et al. Molecular cloning of polymorphic markers on RLGS gel using the spot target cloning method. Biochem. Biophys. Res. Comm. 194, 1406–1412 (1993).

  25. 25

    Hayashizaki, Y. et al. A new method for constructing Noti linking and boundary libraries using a restriction trapper. Genomics 14, 733–739 (1992).

  26. 26

    Hayashizaki, Y. et al. Restriction landmark genomic scanning method and its various applications. Electrophoresis 14, 251–258 (1993).

  27. 27

    Eckert, W.A., Plass, C., Weith, A., Traut, W. & Winking, H. Transcripts from amplified sequences of an inherited homogeneously staining region in chromosome 1 of the house mouse (Mus musculus). Mol. Cell. Biol. 11, 2229–2235 (1991).

  28. 28

    Sambrook, J., Fritsch, E.F. & Maniatis, T. Molecular cloning: A Laboratory Manual, 2nd edn. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989).

  29. 29

    Altschul, S.F., Gish, W., Miller, W., Meyers, E.W. & Lipman, D.J. Basic local alignment search tool. J. Mol. Biol. 215, 403–410 (1990).

  30. 30

    Gardiner-Garden, M. & Frommer, M. CpG islands in vertebrate genomes. J. Mol. Biol. 196, 261–282 (1987).

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Further reading