Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Oct 11;20(19):5189–5195. doi: 10.1093/nar/20.19.5189

A novel murine homeobox gene isolated by a tissue specific PCR cloning strategy.

M J Kern 1, D P Witte 1, M T Valerius 1, B J Aronow 1, S S Potter 1
PMCID: PMC334304  PMID: 1383943

Abstract

We have identified a novel homeobox gene, designated K-2, using a reverse transcription PCR cloning strategy. Sequence analysis reveals that the homeobox of K-2 is 77.6% homologous at the nucleotide level and 97% identical at the amino acid sequence level to another murine gene, S8. Homeodomain sequence comparisons indicate that K-2 and S8 represent a distinct subclass of paired type homeobox genes. Northern blot analysis of RNA from murine embryos and adult tissues identified multiple transcripts that are expressed in a developmentally specific and tissue restricted manner. Alternate splicing of K-2 at the 3-coding region leads to the inclusion of a chain terminating sequence. In addition, the developmental expression pattern of this gene at day 12 of gestation was determined by in situ hybridization. Expression was observed in diverse mesenchymal cells in craniofacial, pericardial, primitive dermal, prevertebral, and genital structures.

Full text

PDF
5189

Images in this article

Selected References

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

  1. Baumgartner S., Bopp D., Burri M., Noll M. Structure of two genes at the gooseberry locus related to the paired gene and their spatial expression during Drosophila embryogenesis. Genes Dev. 1987 Dec;1(10):1247–1267. doi: 10.1101/gad.1.10.1247. [DOI] [PubMed] [Google Scholar]
  2. Bopp D., Burri M., Baumgartner S., Frigerio G., Noll M. Conservation of a large protein domain in the segmentation gene paired and in functionally related genes of Drosophila. Cell. 1986 Dec 26;47(6):1033–1040. doi: 10.1016/0092-8674(86)90818-4. [DOI] [PubMed] [Google Scholar]
  3. Budd P. S., Jackson I. J. What do the regulators regulate? First glimpse downstream. Trends Genet. 1991 Mar;7(3):74–76. doi: 10.1016/0168-9525(91)90273-s. [DOI] [PubMed] [Google Scholar]
  4. Bürglin T. R., Finney M., Coulson A., Ruvkun G. Caenorhabditis elegans has scores of homoeobox-containing genes. Nature. 1989 Sep 21;341(6239):239–243. doi: 10.1038/341239a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Chalepakis G., Fritsch R., Fickenscher H., Deutsch U., Goulding M., Gruss P. The molecular basis of the undulated/Pax-1 mutation. Cell. 1991 Sep 6;66(5):873–884. doi: 10.1016/0092-8674(91)90434-z. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. De Robertis E. M., Oliver G., Wright C. V. Homeobox genes and the vertebrate body plan. Sci Am. 1990 Jul;263(1):46–52. doi: 10.1038/scientificamerican0790-46. [DOI] [PubMed] [Google Scholar]
  9. Epstein D. J., Vekemans M., Gros P. Splotch (Sp2H), a mutation affecting development of the mouse neural tube, shows a deletion within the paired homeodomain of Pax-3. Cell. 1991 Nov 15;67(4):767–774. doi: 10.1016/0092-8674(91)90071-6. [DOI] [PubMed] [Google Scholar]
  10. Finkelstein R., Smouse D., Capaci T. M., Spradling A. C., Perrimon N. The orthodenticle gene encodes a novel homeo domain protein involved in the development of the Drosophila nervous system and ocellar visual structures. Genes Dev. 1990 Sep;4(9):1516–1527. doi: 10.1101/gad.4.9.1516. [DOI] [PubMed] [Google Scholar]
  11. Garcia-Fernàndez J., Baguñ J., Saló E. Planarian homeobox genes: cloning, sequence analysis, and expression. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7338–7342. doi: 10.1073/pnas.88.16.7338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gehring W. J. Homeo boxes in the study of development. Science. 1987 Jun 5;236(4806):1245–1252. doi: 10.1126/science.2884726. [DOI] [PubMed] [Google Scholar]
  13. Gehring W. J. The molecular basis of development. Sci Am. 1985 Oct;253(4):153–162. [PubMed] [Google Scholar]
  14. Goulding M. D., Chalepakis G., Deutsch U., Erselius J. R., Gruss P. Pax-3, a novel murine DNA binding protein expressed during early neurogenesis. EMBO J. 1991 May;10(5):1135–1147. doi: 10.1002/j.1460-2075.1991.tb08054.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hanes S. D., Brent R. A genetic model for interaction of the homeodomain recognition helix with DNA. Science. 1991 Jan 25;251(4992):426–430. doi: 10.1126/science.1671176. [DOI] [PubMed] [Google Scholar]
  16. Hanes S. D., Brent R. DNA specificity of the bicoid activator protein is determined by homeodomain recognition helix residue 9. Cell. 1989 Jun 30;57(7):1275–1283. doi: 10.1016/0092-8674(89)90063-9. [DOI] [PubMed] [Google Scholar]
  17. Hawkins N. C., McGhee J. D. Homeobox containing genes in the nematode Caenorhabditis elegans. Nucleic Acids Res. 1990 Oct 25;18(20):6101–6106. doi: 10.1093/nar/18.20.6101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hayashi S., Scott M. P. What determines the specificity of action of Drosophila homeodomain proteins? Cell. 1990 Nov 30;63(5):883–894. doi: 10.1016/0092-8674(90)90492-w. [DOI] [PubMed] [Google Scholar]
  19. Hill R. E., Favor J., Hogan B. L., Ton C. C., Saunders G. F., Hanson I. M., Prosser J., Jordan T., Hastie N. D., van Heyningen V. Mouse small eye results from mutations in a paired-like homeobox-containing gene. Nature. 1991 Dec 19;354(6354):522–525. doi: 10.1038/354522a0. [DOI] [PubMed] [Google Scholar]
  20. Hoey T., Levine M. Divergent homeo box proteins recognize similar DNA sequences in Drosophila. Nature. 1988 Apr 28;332(6167):858–861. doi: 10.1038/332858a0. [DOI] [PubMed] [Google Scholar]
  21. Jostes B., Walther C., Gruss P. The murine paired box gene, Pax7, is expressed specifically during the development of the nervous and muscular system. Mech Dev. 1990 Dec;33(1):27–37. doi: 10.1016/0925-4773(90)90132-6. [DOI] [PubMed] [Google Scholar]
  22. Kamb A., Weir M., Rudy B., Varmus H., Kenyon C. Identification of genes from pattern formation, tyrosine kinase, and potassium channel families by DNA amplification. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4372–4376. doi: 10.1073/pnas.86.12.4372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kessel M., Gruss P. Murine developmental control genes. Science. 1990 Jul 27;249(4967):374–379. doi: 10.1126/science.1974085. [DOI] [PubMed] [Google Scholar]
  24. Kim Y., Nirenberg M. Drosophila NK-homeobox genes. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7716–7720. doi: 10.1073/pnas.86.20.7716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Krumlauf R., Holland P. W., McVey J. H., Hogan B. L. Developmental and spatial patterns of expression of the mouse homeobox gene, Hox 2.1. Development. 1987 Apr;99(4):603–617. doi: 10.1242/dev.99.4.603. [DOI] [PubMed] [Google Scholar]
  27. Kuziora M. A., McGinnis W. A homeodomain substitution changes the regulatory specificity of the deformed protein in Drosophila embryos. Cell. 1989 Nov 3;59(3):563–571. doi: 10.1016/0092-8674(89)90039-1. [DOI] [PubMed] [Google Scholar]
  28. Laughon A. DNA binding specificity of homeodomains. Biochemistry. 1991 Dec 3;30(48):11357–11367. doi: 10.1021/bi00112a001. [DOI] [PubMed] [Google Scholar]
  29. Monaghan A. P., Davidson D. R., Sime C., Graham E., Baldock R., Bhattacharya S. S., Hill R. E. The Msh-like homeobox genes define domains in the developing vertebrate eye. Development. 1991 Aug;112(4):1053–1061. doi: 10.1242/dev.112.4.1053. [DOI] [PubMed] [Google Scholar]
  30. Murtha M. T., Leckman J. F., Ruddle F. H. Detection of homeobox genes in development and evolution. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10711–10715. doi: 10.1073/pnas.88.23.10711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Opstelten D. J., Vogels R., Robert B., Kalkhoven E., Zwartkruis F., de Laaf L., Destrée O. H., Deschamps J., Lawson K. A., Meijlink F. The mouse homeobox gene, S8, is expressed during embryogenesis predominantly in mesenchyme. Mech Dev. 1991 Mar;34(1):29–41. doi: 10.1016/0925-4773(91)90089-o. [DOI] [PubMed] [Google Scholar]
  32. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rosa F. M. Mix.1, a homeobox mRNA inducible by mesoderm inducers, is expressed mostly in the presumptive endodermal cells of Xenopus embryos. Cell. 1989 Jun 16;57(6):965–974. doi: 10.1016/0092-8674(89)90335-8. [DOI] [PubMed] [Google Scholar]
  34. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  35. Scott M. P., Tamkun J. W., Hartzell G. W., 3rd The structure and function of the homeodomain. Biochim Biophys Acta. 1989 Jul 28;989(1):25–48. doi: 10.1016/0304-419x(89)90033-4. [DOI] [PubMed] [Google Scholar]
  36. Singh G., Kaur S., Stock J. L., Jenkins N. A., Gilbert D. J., Copeland N. G., Potter S. S. Identification of 10 murine homeobox genes. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10706–10710. doi: 10.1073/pnas.88.23.10706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sommer R., Tautz D. Minimal homology requirements for PCR primers. Nucleic Acids Res. 1989 Aug 25;17(16):6749–6749. doi: 10.1093/nar/17.16.6749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ton C. C., Hirvonen H., Miwa H., Weil M. M., Monaghan P., Jordan T., van Heyningen V., Hastie N. D., Meijers-Heijboer H., Drechsler M. Positional cloning and characterization of a paired box- and homeobox-containing gene from the aniridia region. Cell. 1991 Dec 20;67(6):1059–1074. doi: 10.1016/0092-8674(91)90284-6. [DOI] [PubMed] [Google Scholar]
  39. Treisman J., Gönczy P., Vashishtha M., Harris E., Desplan C. A single amino acid can determine the DNA binding specificity of homeodomain proteins. Cell. 1989 Nov 3;59(3):553–562. doi: 10.1016/0092-8674(89)90038-x. [DOI] [PubMed] [Google Scholar]
  40. Walther C., Gruss P. Pax-6, a murine paired box gene, is expressed in the developing CNS. Development. 1991 Dec;113(4):1435–1449. doi: 10.1242/dev.113.4.1435. [DOI] [PubMed] [Google Scholar]
  41. Wedeen C. J., Kostriken R. G., Matsumura I., Weisblat D. A. Evidence for a new family of evolutionarily conserved homeobox genes. Nucleic Acids Res. 1990 Apr 11;18(7):1908–1908. doi: 10.1093/nar/18.7.1908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Witte D. P., Wiginton D. A., Hutton J. J., Aronow B. J. Coordinate developmental regulation of purine catabolic enzyme expression in gastrointestinal and postimplantation reproductive tracts. J Cell Biol. 1991 Oct;115(1):179–190. doi: 10.1083/jcb.115.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zappavigna V., Renucci A., Izpisúa-Belmonte J. C., Urier G., Peschle C., Duboule D. HOX4 genes encode transcription factors with potential auto- and cross-regulatory capacities. EMBO J. 1991 Dec;10(13):4177–4187. doi: 10.1002/j.1460-2075.1991.tb04996.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES