Abstract
TheGNOM gene is required for pattern formation along the main body axis of the embryo in the flowering plantArabidopsis thaliana. Mutations in theGNOM gene alter the asymmetric division of the zygote and interfere with the formation of distinct apical-basal regions in the developing embryo. We have isolated theGNOM gene by positional cloning, characterised its structure and determined the molecular lesions in mutant alleles. Although the predicted 163 kDa GNOM protein has a conserved domain in common with the yeast secretory protein Sec7p, it is most closely related in size and overall similarity to the product of the yeastYEC2 gene, which is not essential for cell viability. Four fully complementinggnom alleles carry missense mutations in conserved regions, seven partially complementing alleles have premature stop codon mutations and two non-complementing alleles have splice-site lesions. Our results suggest that the GNOM protein acts as a complex of identical subunits and that partial complementation may involve low levels of full-length protein generated by inefficient translational read-through.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Achstetter T, Franzusoff A, Field C, Schekman R (1988)SEC7 encodes an unusual, high molecular weight protein required for membrane traffic from the yeast Golgi apparatus. J Biol Chem 263:11711–11717
Altmann T, Felix G, Jessop A, Kauschmann A, Uwer U, Peña-Cortés H, Willmitzer L (1995)Ac/Ds transposon mutagenesis inArabidopsis thaliana: mutant spectrum and frequency ofDs insertion mutants. Mol Gen Genet 247:646–652
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Anderson KV, Jürgens G, Nüsslein-Volhard C (1985) Establishment of dorsal-ventral polarity in theDrosophila embryo: genetic studies on the role of theToll gene product. Cell 42:779–789
Bancroft I, Wolk CP (1988) Pulsed homogeneous orthogonal field gel electrophoresis (PHOGE). Nucleic Acids Res 16:7405–7418
Beier H, Barciszewska M, Sickinger H-D (1984) The molecular basis for the differential translation of TMV RNA in tobacco protoplasts and wheat germ extracts. EMBO J 3:1091–1096
Berleth T, Jürgens G (1993) The role of themonopteros gene in organising the basal body region of theArabidopsis embryo. Development 118:575–587
Bossi L (1983) Context effects: translation of UAG codon by suppressor tRNA is affected by the sequence following UAG in the message. J Mol Biol 164:73–87
Brown JWS (1986) A catalogue of splice junction and putative branch point sequences from plant introns. Nucleic Acids Res 14:9549–9559
Caboche M, Rouzé P (1990) Nitrate reductase: a target for molecular and cellular studies in higher plants. Trends Genet 6:187–192
Chang C, Bowman JL, DeJohn AW, Lander ES, Meyerowitz EM (1988) Restriction fragment length polymorphism linkage map forArabidopsis thaliana. Proc Natl Acad Sci USA 85:6856–6860
Entwistle J, Knudsen S, Müller M, Cameron-Mills V (1991) Amber codon suppression: the in vivo and in vitro analysis of two C-hordein genes from barley. Plant Mol Biol 17:1217–1231
Franzusoff A, Schekman R (1989) Functional compartments of the yeast Golgi apparatus are defined by thesec7 mutation. EMBO J 8:2695–2702
Giraudat J, Hauge BM, Valon C, Smalle J, Parcy F, Goodman HM (1992) Isolation of theArabidopsis ABI3 gene by positional cloning. Plant Cell 4:1251–1261
Grill E, Somerville C (1991) Construction and characterisation of a yeast artificial chromosome library ofArabidopsis which is suitable for chromosome walking. Mol Gen Genet 226:484–490
Haffter P, McMullin TW, Fox TD (1990) A genetic link between an mRNA-specific translational activator and the translation system in yeast mitochondria. Genetics 125:495–503
Hauge BM, Goodman HM (1992) Genome mapping inArabidopsis. In: Koncz C, Chua NH, Schell J (eds) Methods inArabidopsis research. World Scientific Publishers, Singapore, pp 191–223
Ho C-Y, Adamson JG, Hodges RS, Smith M (1994) Heterodimerization of the yeast MATa1 and MATα2 proteins is mediated by two leucine zipper-like coiled-coil motifs. EMBO J 13:1403–1413
Joshi CP (1987) An inspection of the domain between putative TATA box and translational start site in 79 plant genes. Nucleic Acids Res 15:6643–6651
Jürgens G (1995) Axis formation in plant embryogenesis: cues and clues. Cell 81:467–470
Matallana E, Bell CJ, Dunn PJ, Lu M, Ecker JR (1992) Genetical and physical linkage of theArabidopsis genome: methods for anchoring yeast artificial chromosomes. In: Koncz C, Chua NH, Schell J (eds) Methods inArabidopsis research. World Scientific Publishers, Singapore, pp 144–169
Mayer U, Torres Ruiz RA, Berleth T, Miséra S, Jürgens G (1991) Mutations affecting body organization in theArabidopsis embryo. Nature 353:402–407
Mayer U, Büttner G, Jürgens G (1993) Apical-basal pattern formation in theArabidopsis embryo: studies on the role of thegnom gene. Development 117:149–162
Meinke DW (1985) Embryo-lethal mutants ofArabidopsis thaliana: analysis of mutants with a wide range of lethal phases. Theor Appl Genet 69:543–552
Miller JM, Albertini AM (1983) Effects of surrounding sequence on the suppression of nonsense codons. J Mol Biol 164:59–71
Olszewski NE, Martin FB, Ausubel FM (1988) Specialized binary vector for plant transformation: expression of theArabidopsis thaliana AHAS gene inNicotiana tabacum. Nucleic Acids Res 16:10765–10782
Pruitt RE, Meyerowitz EM (1986) Characterisation of the genome ofArabidopsis thaliana. J Mol Biol 187:169–183
Putterill J, Robson F, Lee K, Coupland G (1993) Chromosome walking with YAC clones inArabidopsis: isolation of 1700 kb of contiguous DNA on chromosome 5, including a 300 kb region containing the flowering-time geneCO. Mol Gen Genet 239:145–157
Riley J, Butler R, Ogilvie D, Finniear R, Jenner D, Powell S, Anand R, Smith JC, Markham AF (1990) A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones. Nucleic Acids Res 18:2887–2890
Rogers SO, Bendich AJ (1988) Extraction of DNA from plant tissue. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual A6. Kluwer Academic Publishers, Dordrecht, pp 1–10
Rose MD, Winston F, Hieter P (1988) Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning (2nd edn) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schneider DS, Hudson KL, Lin T-Y, Anderson KV (1991) Dominant and recessive mutations define functional domains ofToll, a transmembrane protein required for dorsal-ventral polarity in theDrosophila embryo. Genes Dev 5:797–807
Shevell DE, Leu WM, Gillmor CS, Xia G, Feldmann KA, Chua N-H (1994)EMB30 is essential for normal cell division, cell expansion, and cell adhesion inArabidopsis and encodes a protein that has similarity to Sec7. Cell 77:1051–1062
Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA inSaccharomyces cerevisiae. Genetics 125:19–27
Ward ER, Jen GC (1990) Isolation of single-copy-sequence clones from a yeast artificial chromosome library of randomly shearedArabidopsis thaliana DNA. Plant Mol Biol 14:561–568
Weigel D, Alvarez J, Smyth DR, Yanofsky MF, Meyerowitz EM (1992)LEAFY controls floral meristem identity inArabidopsis. Cell 69:843–859
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Busch, M., Mayer, U. & Jürgens, G. Molecular analysis of theArabidopsis pattern formation geneGNOM: gene structure and intragenic complementation. Molec. Gen. Genet. 250, 681–691 (1996). https://doi.org/10.1007/BF02172979
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02172979