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First published online 18 July 2007
doi: 10.1242/dev.009373

Development 134, 2991-2999 (2007)
Published by The Company of Biologists 2007
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Two C. elegans histone methyltransferases repress lin-3 EGF transcription to inhibit vulval development

Erik C. Andersen and H. Robert Horvitz*

Howard Hughes Medical Institute, Department of Biology, MIT, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.


Figure 1
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  		Fig. 1. met-1 and met-2 gene structures, mutations and predicted protein structures. (A) The genomic structures of C. elegans met-1 and met-2. Exons are represented by black boxes, 3' untranslated regions by white boxes. The alternatively spliced exon six of met-1 is depicted as a white, stippled box. The locations of the deletion alleles are shown. (B) The domain structures of the MET-1 and MET-2 proteins.

 

Figure 2
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  		Fig. 2. met-1 and met-2 are required in vivo for normal levels of histone H3K36 and H3K9 trimethylation, respectively. The levels of histone H3 trimethylation at K4, K9, K27 and K36 were assayed using quantitative western blots and normalized to levels of histone H3 (see Materials and methods). Relative histone H3 trimethylation levels of met-1(n4337) mutants (white) and met-2(n4256) mutants (gray) were normalized to the trimethylation levels of wild-type C. elegans (black) for each experiment. The levels of histone H3 were measured (see Fig. S1 in the supplementary material), and the specificity of the H3K9 and H3K36 trimethylation antisera were confirmed using dot blots of methylated histone tail peptides (see Fig. S2 in the supplementary material). Normalized units of fluorescence and standard deviations are shown.

 

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  		Fig. 3. met-1 and met-2 are each required to prevent ectopic lin-3 expression in a lin-15A(n767) mutant background. Real-time RT-PCR experiments were performed using RNA samples from C. elegans of the genotypes shown. Mean {Delta}{Delta}CT values were used to calculate relative changes in lin-3 expression normalized to levels of rpl-26 (see Materials and methods). Mean values and ranges of relative lin-3/rpl-26 ratios for three trials are shown.

 




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