Hypermethylated SUPERMAN epigenetic alleles in Arabidopsis

Science

Volumn 277, Issue 5329, 1997, Pages 1100-1103

  Jacobsen, Steven E ^a   Meyerowitz, Elliot M ^a  

^a CALIFORNIA INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALLELE; ARABIDOPSIS; ARTICLE; COMPLEX FORMATION; DNA METHYLATION; GENE DUPLICATION; GENE MUTATION; GENOME IMPRINTING; MEMORY CELL; METHYLATION; NONHUMAN; PRIORITY JOURNAL; X CHROMOSOME;

ALLELES; ARABIDOPSIS; ARABIDOPSIS PROTEINS; BASE SEQUENCE; CROSSES, GENETIC; CYTOSINE; DNA (CYTOSINE-5-)-METHYLTRANSFERASE; DNA METHYLATION; DNA, ANTISENSE; DNA, PLANT; GENE EXPRESSION REGULATION, PLANT; GENES, PLANT; GENETIC COMPLEMENTATION TEST; MOLECULAR SEQUENCE DATA; MUTATION; PHENOTYPE; PLANTS, GENETICALLY MODIFIED; RNA, MESSENGER; RNA, PLANT; TRANSCRIPTION FACTORS;

ARABIDOPSIS;

EID: 0030769859     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5329.1100     Document Type: Article

References (23)

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4. E. J. Finnegan, W. J. Peacock, E. S. Dennis, Proc. Natl. Acad. Sci. U.S.A. 93, 8449 (1996).
5. M. J. Ronemus, M. Galbiati, C. Ticknor, J. Chen, S. L. Dellaporta, Science 273, 654 (1996).
6. clk-2 and clk-3 were from an ethylmethanesulfonate mutagenesis, clk-5 from a diepoxybutane mutagenesis, and clk-6 from a transferred-DNA insertional mutagenesis. clk-1, clk-4, and clk-7 were from the fwa-7, gl2-1, and tt2-1 mutants, respectively, obtained from the Arabidopsis Biological Resource Center. All alleles are in the Ler background.
7. J. L. Bowman et al., Development 114, 599 (1992); E. A. Schultz, F. B. Pickett, G. W. Haughn, Plant Cell 3, 1221 (1991).
8. J. L. Bowman et al., Development 114, 599 (1992); E. A. Schultz, F. B. Pickett, G. W. Haughn, Plant Cell 3, 1221 (1991).
9. J. C. Gaiser, K. Robinson-Beers, C. S. Gasser, Plant Cell 7, 333 (1995).
10. H. Sakai and E. M. Meyerowitz, personal communication.
11. Wild-type plants usually have two carpels per flower, but occasionally have extra carpels. clk and sup heterozygotes contain more extra carpels than the wild type. Whereas Ler plants had 0.2 ± 0.2 extra carpels per plant (mean ± SE) in the first 10 flowers, clk-3 heterozygotes had 3.3 ± 0.7, and sup-5 heterozygotes had 2.1 ± 0.7.
12. C. Chang, J. L. Bowman, A. W. DeJohn, E. Lander, E. M. Meyerowitz, Proc. Natl. Acad. Sci. U.S.A. 85, 6856 (1988).
13. H. Sakai, L. J. Medrano, E. M. Meyerowitz, Nature 378, 199 (1995).
14. Genomic DNA from Ler or clk-3 plants was partially digested with Sau 3A and cloned in the LambdaGEM-11 vector (Promega). The adjacent 5.5-and 1.2-kb Eco Rl SUP genomic fragments (11) were subcloned from hybridization-selected lambda clones into the plant transformation vector pCGN1547 [K. E. McBride and K. R. Summerfelt, Plant Mol. Biol. 14, 269 (1990)] and then transformed into either clk-3 or sup-5 plants [N. Bechtold, J. Ellis, G. Pelletier, C. R. Acad. Sci. Paris 316, 1194 (1993)].
15. Genomic DNA from Ler or clk-3 plants was partially digested with Sau 3A and cloned in the LambdaGEM-11 vector (Promega). The adjacent 5.5-and 1.2-kb Eco Rl SUP genomic fragments (11) were subcloned from hybridization-selected lambda clones into the plant transformation vector pCGN1547 [K. E. McBride and K. R. Summerfelt, Plant Mol. Biol. 14, 269 (1990)] and then transformed into either clk-3 or sup-5 plants [N. Bechtold, J. Ellis, G. Pelletier, C. R. Acad. Sci. Paris 316, 1194 (1993)].
16. CosTH1 was isolated from a Wassilewskija genomic cosmid library (Arabidopsis Biological Resource Center) and contains a 25-kb insert that partially overlaps the proximal side of the SUP 6.7-kb genomic region. An overlapping set of cosmid clones on the distal side of SUP (LM5-2, LM5-8, Q8PMl, Pst14.5, Pst11, and Bam5.5) was a gift from H. Sakai.
17. Recently, three mutants were described [H. Huang and H. Ma, Plant Cell 9, 115 (1997)] that also have a weak sup-like phenotype. Although the authors interpreted these mutations as being in a previously unidentified gene near SUP, it seems possible that these are also epigenetic SUP alleles.
18. M. Koornneef, S. W. M. Dellaert, J. H. van der Veen, Mutat. Res. 93, 109 (1982).
19. S. E. Jacobsen and E. M. Meyerowitz, data not shown.
20. Genomic DNA from whole shoots was used for bisulfite sequencing [R. Feil, J. Charlton, A. P. Bird, J. Walter, W. Reik, Nucleic Acids Res. 22, 695 (1994)]. Seven overlapping polymerase chain reaction (PCR) amplifications were used to determine the methylation pattern shown in Fig. 3. PCR products were sequenced with Amplitaq DNA Polymerase FS dye terminator cycle sequencing (Perkin-Elmer).
21. The SUP cDNA was used to probe genomic DNA blots at various stringencies. Under high-stringency conditions (10), only the SUP gene is detected. At lower washing stringency (2x saline sodium phosphate EDTA, 0.5% SDS, 55°C), an additional band is detected. This hybridizing fragment was cloned and found to contain a region of homology limited to a 101-bp sequence, encoding a zinc-finger domain, which shares 76% nucleic acid identity with SUP.
22. D. R. Smyth, J. L. Bowman, E. M. Meyerowitz, Plant Cell 2, 755 (1990).
23. We thank G. Serraiocco for technical assistance, H. Sakai for SUP genomic sequences and cosmid clones, E. Finnegan and E. Dennis for the AMT line, D. Weigel for clk-5, J. Viret and E. Signer for clk-6, and X. Chen, J. Fletcher, M. Frohlich, J. Hua, C. Ohno, J. L. Riechmann, R. Sablowski, H. Sakai, D. Wagner, and E. Ziegelhoffer for review of the manuscript. S.E.J. was supported by an NIH postdoctoral fellowship. This work was supported by NSF grant MCB-9204839 to E.M.M.