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First published online 19 July 2006
doi: 10.1242/dev.02490

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Promotion of oogenesis and embryogenesis in the C. elegans gonad by EFL-1/DPL-1 (E2F) does not require LIN-35 (pRB)

Department of Genetics, Yale University School of Medicine, New Haven, CT 06520, USA.

View larger version (29K): [in a new window]   Fig. 1. C. elegans germline. The right arm of the gonad shows germ cell development. The left arm of the gonad delineates the distal, medial and proximal regions, as referred to in the text. Fertilized embryos are present in the uterus. Orange, proliferation; blue, meiotic prophase I; yellow, maturing oocytes; red, sperm; brackets, region in which EFL-1 protein is detectably expressed.

View larger version (29K): [in a new window]   Fig. 2. Dissected gonads from efl-1, dpl-1 and lin-35 mutants. Hermaphrodite gonads from (A) wild type (N2); (B) lin-35(n745); (C) dpl-1(n3316); and (D) efl-1(n3639). Distal end of gonad marked with asterisk. Scale bars: 50 µm. Dia, oocyte in diakinesis of meiosis I; Emo, endomitotic.

View larger version (24K): [in a new window]   Fig. 3. Target genes of lin-35, efl-1 and dpl-1. The overlap between genes regulated at least twofold, P<0.05 in at least one mutant are displayed by Venn diagram. Genes are considered overlapping if they are regulated >1.5x, P<0.05 in the second mutant. Regulated genes are divided into downregulated in mutants (A) and upregulated in mutants (B). Three genes (col-178, asp-1 and F11A3.2) are represented twice, as they show opposite regulation in two mutants (see Table S1 in the supplementary material).

View larger version (30K): [in a new window]   Fig. 4. Hierarchical cluster analysis of lin-35-, efl-1- and dpl-1-regulated genes. The 272 genes are displayed in rows; columns represent the average of repeats for each mutant (D, dpl-1; E, efl-1; L, lin-35). Red, lower expression in mutant samples relative to controls; black, equivalent expression in mutant relative to control; green, higher expression in mutant relative to control samples.

View larger version (22K): [in a new window]   Fig. 5. Identification of candidate regulatory motifs upstream of target genes. (A,C) Graphic representation of motifs identified through MEME. The height of a letter represents the relative frequency of occurrence. (B,D) Location of putative motifs within a 1 kb region upstream of selected candidates. Small boxes indicate the location of the motif. Scale at bottom in nucleotides.

View larger version (102K): [in a new window]   Fig. 6. In-situ hybridization patterns of example Group I and Group IV genes. The head of the animal is marked by an asterisk. (A-D) Autosomal Group I genes initiate detectable expression in the medial germline, while X-linked genes are more proximal. Arrows mark medial germline and onset of expression. (E-H) Group IV genes generally show broad expression, including distal and proximal germline. In-situ data courtesy of Y. Kohara (http://nematode.lab.nig.ac.jp).

View larger version (79K): [in a new window]   Fig. 7. Reduction of RME-2 and MEX-5 levels in efl-1(n3363) and dpl-1(n3316) mutants. (A-L) Adult germlines from each genotype were dissected, fixed and stained with an antibody to the corresponding protein as labeled. Blue, DNA; red, antibody. Panels G and J are at 10x exposure relative to A. For each antibody and genotype, n>20. The reduced RME-2 and MEX-5 staining seen in efl-1 and dpl-1 mutants occurred in all gonads examined. LIN-3 staining on the oocyte surface disappears upon RNAi of lin-3, whereas the diffuse staining does not (see Fig. S2 in the supplementary material).