Summary
Paramutation is observed when the Antirrhinum majus lines 44 and 53 are crossed. These two lines both have insertions at the nivea locus, which encodes chalcone synthase (chs). The allele niv-53 carries the transposable element Tam1 in the promoter region of the chs gene; niv-44 carries the element Tam2 within the gene. The Tam1 element has previously been extensively characterised. Here the Tam2 element is further characterised, and the arrangement of the nivea locus in paramutant plants is analysed. The complete sequence of Tam2, and that of a partial cDNA complementary to it, have been determined. The cDNA is probably transcribed from a different copy of Tam2 from that present at the nivea locus, and does not encode a functional protein. Genomic Southerns of F1 plants from the 53/44 cross show that no major rearrangements are consistently associated with paramutation at the nivea locus of A. majus. The isolation from a paramutant plant arising from a 53/44 cross of an allele (niv-4432) resulting from the excision of Tam2 is reported. The excision of Tam2 resulted in a 32 bp deletion of chs gene sequences. Plants homozygous for the new niv-4432 allele have white flowers and are still paramutagenic, demonstrating that Tam2 need not be present at the nivea locus for paramutation to occur. Different interactions between Tam1 and Tam2 are discussed, and a possible model for paramutation is presented.
Similar content being viewed by others
References
Benton WD, Davis RW (1977) Screening lambda gt recombinant clones by hybridization to single plaques in situ. Science 196:180–182
Bonas U, Sommer H, Harrison BJ, Saedler H (1984a) The transposable element Tam1 of Antirrhinum majus is 17 kb long. Mol Gen Genet 194:138–143
Bonas U, Sommer H, Saedler H (1984b) The 17-kb Tam1 element of Antirrhinum majus induces a 3-bp duplication upon integration into the chalcone synthase gene. EMBO J 3:1015–1019
Breathnach R, Chambon P (1981) Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem 50:349–383
Brink RA (1973) Paramutation. Annu Rev Genet 7:129–152
Brink RA, Styles ED, Axtell JD (1968) Paramutation: Directed genetic change. Science 159:161–170
Brown DF, Brink RA (1960) Paramutagenic action of paramutant Rr and Rg alleles in maize. Genetics 45:1313–1316
Burd FJ, Wells RD (1974) Synthesis and characterization of the duplex block polymer d(C15A15).(T15G15). J Biol Chem 249:7094–7101
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Döring H-P, Starlinger P (1984) Barbara McClintock's controlling elements: Now at the DNA level. Cell 39:253–259
Englund PT (1971) Analysis of nucleotide sequences at 3′ termini of duplex deoxyribonucleic acid with the use of the T4 deoxyribonucleic acid polymerase. J Biol Chem 246:3269–3276
Gubler U, Hoffman BJ (1983) A simple and very efficient method for generating cDNA libraries. Gene 25:263–269
Hagemann R, Berg W (1977) Vergleichende Analyse der Paramutationssysteme bei höheren Pflanzen. Biol Zentralblatt 96:257–301
Harrison BJ, Carpenter R (1973) A comparison of the instabilities at the nivea and pallida loci in Antirrhinum majus. Heredity 31:309–323
Hehl R, Sommer H, Saedler H (1987) Interaction between the Tam1 and Tam2 transposable elements of Antirrhinum majus. Mol Gen Genet 207:47–53
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Hohn B (1979) In vitro packaging of lambda and cosmid DNA. Methods Enzymol 68:299–309
Hudson A, Carpenter R, Coen ES (1987) De novo activation of the transposable element Tam2 of Antirrhinum majus. Mol Gen Genet 207:54–59
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Meinkoth J, Wahl G (1984) Hybridization of nucleic acids on solid supports. Anal Biochem 138:267–284
Murray NE (1983) Phage lambda and molecular cloning. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory Press, New York, pp 395–432
Nevers P, Shepherd NS, Saedler H (1986) Plant transposable elements. Adv Bot Res 12:103–203
Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligodeoxyribonucleotide-directed mutagenesis. Gene 26:101–106
Pereira A, Schwarz-Sommer Zs, Gierl A, Bertram I, Peterson PA, Saedler H (1985) Genetic and molecular analysis of the Enhancer (En) transposable element system of Zea mays. EMBO J 4:17–23
Reed R, Maniatis T (1986) A role for exon sequences and splice-site proximity in splice-site selection. Cell 46:681–690
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schwarz-Sommer Zs, Gierl A, Kloesgen RB, Wienand U, Peterson PA, Saedler H (1984) The Spm (En) transposable element controls the excision of a 2-kb DNA insert at the wxm-8 allele of Zea mays. EMBO J 3:1021–1028
Schwarz-Sommer Zs, Shepherd N, Tacke E, Gierl A, Rohde W, Leclercq L, Mattes M, Berndtgen R, Peterson PA, Saedler H (1987) Influence of transposable elements on the structure and function of the A1 gene of Zea mays. EMBO J 6:287–294
Sommer H, Saedler H (1986) Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 202:429–434
Spribille R, Forkmann G (1982) Genetic control of chalcone synthase activity in flowers of Antirrhinum majus. Phytochem 21:2231–2234
Stüber K (1986) Nucleic acid secondary structure prediction and display. Nucleic Acids Res 14:317–326
Sutton WD, Gerlach WL, Schwartz D, Peacock WJ (1984) Molecular analysis of Ds controlling element mutations at the AdhI locus of maize. Science 223:1265–1268
Taylor B, Powell A (1982) Isolation of plant DNA and RNA. Bethesda Research Laboratory Focus 4:4–6
Upadhyaya KC, Sommer H, Krebbers E, Saedler H (1985) The paramutagenic line niv-44 has a 5 kb insert, Tam2, in the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 199:201–207
Wartell RM, Reznikoff WS (1980) Cloning DNA restriction endonuclease fragments with protruding single-stranded ends. Gene 9:307–319
Wienand U, Schwarz Zs, Feix G (1979) Electrophoretic elution of nucleic acids from gels adapted for subsequent biological tests. FEBS Lett 98:319–323
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Krebbers, E., Hehl, R., Piotrowiak, R. et al. Molecular analysis of paramutant plants of Antirrhinum majus and the involvement of transposable elements. Mol Gen Genet 209, 499–507 (1987). https://doi.org/10.1007/BF00331156
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331156